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strain atcc 3624  (ATCC)


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    Structured Review

    ATCC strain atcc 3624
    Strain Atcc 3624, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/atcc+3624/Clostridium+perfringens%3B+Strain+26/pm40301878-174-11-12
    Average 93 stars, based on 45 article reviews
    strain atcc 3624 - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Produced:

    Article Title: The Agr-Like Quorum-Sensing System Is Important for Clostridium perfringens Type A Strain ATCC 3624 To Cause Gas Gangrene in a Mouse Model
    Article Snippet: .. Consistent with previous observations (25, 30) using type A strain 13, inactivation of the agrB gene in ATCC 3624 also produced a significant in vitro decrease in cpa and pfoA expression at the transcriptional level and production of these protein toxins at the translational level. ..

    In Vitro:

    Article Title: The Agr-Like Quorum-Sensing System Is Important for Clostridium perfringens Type A Strain ATCC 3624 To Cause Gas Gangrene in a Mouse Model
    Article Snippet: .. Consistent with previous observations (25, 30) using type A strain 13, inactivation of the agrB gene in ATCC 3624 also produced a significant in vitro decrease in cpa and pfoA expression at the transcriptional level and production of these protein toxins at the translational level. ..

    Article Title: The Agr-Like Quorum-Sensing System Is Important for Clostridium perfringens Type A Strain ATCC 3624 To Cause Gas Gangrene in a Mouse Model
    Article Snippet: .. Effects of the 6-R synthetic signaling peptide on in vitro CPA or PFO production by ATCC 3624. ..

    Expressing:

    Article Title: The Agr-Like Quorum-Sensing System Is Important for Clostridium perfringens Type A Strain ATCC 3624 To Cause Gas Gangrene in a Mouse Model
    Article Snippet: .. Consistent with previous observations (25, 30) using type A strain 13, inactivation of the agrB gene in ATCC 3624 also produced a significant in vitro decrease in cpa and pfoA expression at the transcriptional level and production of these protein toxins at the translational level. ..

    Article Title: The presence of differentiated C2C12 muscle cells enhances toxin production and growth by Clostridium perfringens type A strain ATCC3624
    Article Snippet: .. The error bars indicate the S.D. (a) RT-qPCR (5 ng of cDNA) comparing agrB , agrD and virS expression by ATCC 3624 cultured in the presence or absence or differentiated C2C12 cells. ..

    other:

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21
    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.


    Thin Layer Chromatography:

    Article Title: Lysyl-Phosphatidylglycerol: A Lipid Involved in the Resistance of Staphylococcus aureus to Antimicrobial Peptide Activity
    Article Snippet: Caulobacter acrescentes , Gram-negative , CB13 , 10.4–11.4% , Phospholipids were analyzed from extracted 32 P-labeled lipids located in the chromatogram by autoradiography. , [ ] . .. Clostridium perfringens , Gram-positive , Is not named ATCC 3624 , + + , TLC and infrared spectra TLC and MS , [ ] [ ] . ..

    Quantitative RT-PCR:

    Article Title: The presence of differentiated C2C12 muscle cells enhances toxin production and growth by Clostridium perfringens type A strain ATCC3624
    Article Snippet: .. The error bars indicate the S.D. (a) RT-qPCR (5 ng of cDNA) comparing agrB , agrD and virS expression by ATCC 3624 cultured in the presence or absence or differentiated C2C12 cells. ..

    Cell Culture:

    Article Title: The presence of differentiated C2C12 muscle cells enhances toxin production and growth by Clostridium perfringens type A strain ATCC3624
    Article Snippet: .. The error bars indicate the S.D. (a) RT-qPCR (5 ng of cDNA) comparing agrB , agrD and virS expression by ATCC 3624 cultured in the presence or absence or differentiated C2C12 cells. ..



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    Figure 1. Expression of brnQ genes during culture in TY cultures of <t>ATCC3624.</t> (A) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expres- sion), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in Table 1. The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. (B) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.
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    Expression of brnQ genes during culture in TY cultures of <t>ATCC3624.</t> ( A ) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expression), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in . The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. ( B ) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.
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    Expression of brnQ genes during culture in TY cultures of <t>ATCC3624.</t> ( A ) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expression), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in . The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. ( B ) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.
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    Expression of brnQ genes during culture in TY cultures of <t>ATCC3624.</t> ( A ) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expression), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in . The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. ( B ) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.
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    Figure 1. Expression of brnQ genes during culture in TY cultures of ATCC3624. (A) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expres- sion), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in Table 1. The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. (B) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 1. Expression of brnQ genes during culture in TY cultures of ATCC3624. (A) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expres- sion), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in Table 1. The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. (B) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Positive Control, Negative Control, Comparison, Software

    Figure 2. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ1 null mutant (brnQ1KO), and the complementing strain (brnQ1c) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures of these strains from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC level in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the sample from the same 3 h TY sample, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ1 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The graph shows the mean values from three independent experiments. The error bars indicate the S.D. * p < 0.05 relative to wild type. Similar Image J analyses did not detect PLC production differences among these strains in 7 or 24 h samples (Figure S5A). (D) Western blot analyses of the PFO level in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points (Figure S5A).

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 2. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ1 null mutant (brnQ1KO), and the complementing strain (brnQ1c) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures of these strains from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC level in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the sample from the same 3 h TY sample, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ1 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The graph shows the mean values from three independent experiments. The error bars indicate the S.D. * p < 0.05 relative to wild type. Similar Image J analyses did not detect PLC production differences among these strains in 7 or 24 h samples (Figure S5A). (D) Western blot analyses of the PFO level in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points (Figure S5A).

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    Figure 3. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ2 null mutant (brnQ2KO), and the complementing strain (brnQ2c) when cultured in TY medium. (A) Post-inoculation changes in the OD600 of TY cultures of these strains from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) The OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the same 3 h TY samples, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ2 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among the strains in 7 or 24 h samples (Figure S5B). (D) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown at left. Image J software did not detect significant differences in PFO production among the strains at any time points (Figure S5B).

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 3. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ2 null mutant (brnQ2KO), and the complementing strain (brnQ2c) when cultured in TY medium. (A) Post-inoculation changes in the OD600 of TY cultures of these strains from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) The OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the same 3 h TY samples, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ2 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among the strains in 7 or 24 h samples (Figure S5B). (D) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown at left. Image J software did not detect significant differences in PFO production among the strains at any time points (Figure S5B).

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    Figure 4. A comparison of the post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ3 null mutant (brnQ3KO), and the complementing strain (brnQ3c) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) The OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among these strains in 7 or 24 h samples (Figure S5C). (D) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points (Figure S5C).

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 4. A comparison of the post-inoculation changes in culture OD600, vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ3 null mutant (brnQ3KO), and the complementing strain (brnQ3c) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures from 0 to 9 h at 37 ◦C. The result shown is representative of three repetitions. (B) The OD600 of TY cultures at 24 h at 37 ◦C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among these strains in 7 or 24 h samples (Figure S5C). (D) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points (Figure S5C).

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    Figure 5. A comparison of expression of the brnQ genes by wild-type ATCC3624, its isogenic brnQ null mutants, and complementing strains. ATCC3624, the single brnQ null mutants, or complementing strains were inoculated into TY broth and then incubated anaerobically for 2 h at 37 ◦C. Bacteria were collected and pelleted via centrifugation. Total RNA was extracted from the pellets, and cDNA was made for RT-qPCR analyses. Average CT values were normalized to the housekeeping recA gene, and the fold differences in expression were calculated using the comparative CT method (2−∆∆CT). The mean values from three independent experiments are shown. The error bars indicate the S.D. (A) RT- qPCR for the expression of brnQ2 or brnQ3 was performed using cDNA from the wild-type parent, the brnQ1 null mutant, and the complementing strain after their incubation in a TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. (B) RT-qPCR for the expression of brnQ1 or brnQ3 was performed using cDNA from the wild type-parent, the brnQ2 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. (C) RT-qPCR for the expression of brnQ1 or brnQ2 was performed using cDNA from the wild-type parent, the brnQ3 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 5. A comparison of expression of the brnQ genes by wild-type ATCC3624, its isogenic brnQ null mutants, and complementing strains. ATCC3624, the single brnQ null mutants, or complementing strains were inoculated into TY broth and then incubated anaerobically for 2 h at 37 ◦C. Bacteria were collected and pelleted via centrifugation. Total RNA was extracted from the pellets, and cDNA was made for RT-qPCR analyses. Average CT values were normalized to the housekeeping recA gene, and the fold differences in expression were calculated using the comparative CT method (2−∆∆CT). The mean values from three independent experiments are shown. The error bars indicate the S.D. (A) RT- qPCR for the expression of brnQ2 or brnQ3 was performed using cDNA from the wild-type parent, the brnQ1 null mutant, and the complementing strain after their incubation in a TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. (B) RT-qPCR for the expression of brnQ1 or brnQ3 was performed using cDNA from the wild type-parent, the brnQ2 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. (C) RT-qPCR for the expression of brnQ1 or brnQ2 was performed using cDNA from the wild-type parent, the brnQ3 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Expressing, Incubation, Bacteria, Centrifugation, Quantitative RT-PCR, Mutagenesis

    Figure 7. A comparison of PLC or PFO production levels for wild-type ATCC3624, its double null mutants, and the complementing strain when cultured in a TY medium. Western blot analyses of PLC (A) and PFO (B) levels in supernatants of the wild type, double null mutants, and their complementing strains in the early-stage culture (3 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right figure in panels A and B show the comparison of Western blot intensities determined by Image J. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 7. A comparison of PLC or PFO production levels for wild-type ATCC3624, its double null mutants, and the complementing strain when cultured in a TY medium. Western blot analyses of PLC (A) and PFO (B) levels in supernatants of the wild type, double null mutants, and their complementing strains in the early-stage culture (3 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right figure in panels A and B show the comparison of Western blot intensities determined by Image J. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Cell Culture, Western Blot

    Figure 8. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, or PLC and PFO production levels for ATCC3624 (WT) vs. the triple null mutant (TKO) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures incubated for up to 9 h at 37 ◦C. The results shown are representative of three repetitions. (B) The OD600 of TY cultures after 24 h of incubation at 37 ◦C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio between 3 h samples (right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PLC production differences among these strains in 7 or 24 h samples (Figure S10A). (D) Western blot analyses of PFO levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right graph in panel D shows the comparison of the Western blot band intensity ratio between 3 h samples using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PFO production differences among these strains in 7 or 24 h samples (Figure S10B).

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624.

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Figure 8. A comparison of post-inoculation changes in culture OD600, vegetative cell viability, or PLC and PFO production levels for ATCC3624 (WT) vs. the triple null mutant (TKO) when cultured in a TY medium. (A) Post-inoculation changes in the OD600 of TY cultures incubated for up to 9 h at 37 ◦C. The results shown are representative of three repetitions. (B) The OD600 of TY cultures after 24 h of incubation at 37 ◦C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. (C) Western blot analyses of PLC levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio between 3 h samples (right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PLC production differences among these strains in 7 or 24 h samples (Figure S10A). (D) Western blot analyses of PFO levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right graph in panel D shows the comparison of the Western blot band intensity ratio between 3 h samples using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PFO production differences among these strains in 7 or 24 h samples (Figure S10B).

    Article Snippet: The current study used C. perfringens type A strain ATCC3624 (ATCC®, Manassas VA, USA) and E. coli DH5α competent cells [New England Biolabs (NEB), Ipswich, MA, USA].

    Techniques: Comparison, Mutagenesis, Cell Culture, Incubation, Western Blot, Software

    Expression of brnQ genes during culture in TY cultures of ATCC3624. ( A ) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expression), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in . The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. ( B ) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: Expression of brnQ genes during culture in TY cultures of ATCC3624. ( A ) RT-PCR analyses. The recA gene was used as a housekeeping gene, and a sample lacking reverse transcriptase (RT-) was run (top panel) in a PCR reaction with recA primers to show the absence of DNA contamination in these RNA samples; recA gene RT-PCR (2nd panel) showed that cDNA was present in all samples. Further RT-PCR analyses (bottom three panels) are shown using brnQ1 primers (for brnQ1 expression), brnQ2 primers (for brnQ2 expression), or brnQ3 primers (for brnQ3 expression) for wild-type ATCC3624 cultures at different time points. The expected size of each PCR product is listed in . The sample with ATCC3624 DNA served as the positive control, and the sample without ATCC3624 DNA served as the negative control. DNA size markers are shown on the left of each gel. The results shown are representative of three repetitions. ( B ) A comparison of the RT-PCR band intensity ratio between the brnQ gene vs. the housekeeping recA gene is shown using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the 2 h result.

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Positive Control, Negative Control, Comparison, Software

    A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ1 null mutant (brnQ1KO), and the complementing strain (brnQ1c) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures of these strains from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC level in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the sample from the same 3 h TY sample, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ1 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The graph shows the mean values from three independent experiments. The error bars indicate the S.D. * p < 0.05 relative to wild type. Similar Image J analyses did not detect PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of the PFO level in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points .

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ1 null mutant (brnQ1KO), and the complementing strain (brnQ1c) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures of these strains from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC level in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the sample from the same 3 h TY sample, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ1 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The graph shows the mean values from three independent experiments. The error bars indicate the S.D. * p < 0.05 relative to wild type. Similar Image J analyses did not detect PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of the PFO level in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points .

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ2 null mutant (brnQ2KO), and the complementing strain (brnQ2c) when cultured in TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures of these strains from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) The OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the same 3 h TY samples, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ2 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among the strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown at left. Image J software did not detect significant differences in PFO production among the strains at any time points .

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ2 null mutant (brnQ2KO), and the complementing strain (brnQ2c) when cultured in TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures of these strains from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) The OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The PLC blot was also repeated using 10 µL more of the same 3 h TY samples, with longer film exposure (bottom left blot), to allow for a better comparison of PLC production between the brnQ2 null mutant vs. wild-type ATCC3624 or the complementing strain. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among the strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown at left. Image J software did not detect significant differences in PFO production among the strains at any time points .

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    A comparison of the post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ3 null mutant (brnQ3KO), and the complementing strain (brnQ3c) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) The OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points .

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of the post-inoculation changes in culture OD 600 , vegetative cell viability, and PLC or PFO production levels for ATCC3624 (WT), its brnQ3 null mutant (brnQ3KO), and the complementing strain (brnQ3c) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures from 0 to 9 h at 37 °C. The result shown is representative of three repetitions. ( B ) The OD 600 of TY cultures at 24 h at 37 °C (top). The same samples were also used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in supernatants of 3, 7, and 24 h culture samples (top panel). The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio is shown between 3 h samples (bottom graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in supernatants of 3, 7, and 24 h culture samples. The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. Image J analyses did not detect significant differences in PFO production among these strains at any time points .

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Mutagenesis, Cell Culture, Western Blot, Software

    A comparison of expression of the brnQ genes by wild-type ATCC3624, its isogenic brnQ null mutants, and complementing strains. ATCC3624, the single brnQ null mutants, or complementing strains were inoculated into TY broth and then incubated anaerobically for 2 h at 37 °C. Bacteria were collected and pelleted via centrifugation. Total RNA was extracted from the pellets, and cDNA was made for RT-qPCR analyses. Average C T values were normalized to the housekeeping recA gene, and the fold differences in expression were calculated using the comparative C T method (2 −ΔΔC T ). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( A ) RT-qPCR for the expression of brnQ2 or brnQ3 was performed using cDNA from the wild-type parent, the brnQ1 null mutant, and the complementing strain after their incubation in a TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. ( B ) RT-qPCR for the expression of brnQ1 or brnQ3 was performed using cDNA from the wild type-parent, the brnQ2 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. ( C ) RT-qPCR for the expression of brnQ1 or brnQ2 was performed using cDNA from the wild-type parent, the brnQ3 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of expression of the brnQ genes by wild-type ATCC3624, its isogenic brnQ null mutants, and complementing strains. ATCC3624, the single brnQ null mutants, or complementing strains were inoculated into TY broth and then incubated anaerobically for 2 h at 37 °C. Bacteria were collected and pelleted via centrifugation. Total RNA was extracted from the pellets, and cDNA was made for RT-qPCR analyses. Average C T values were normalized to the housekeeping recA gene, and the fold differences in expression were calculated using the comparative C T method (2 −ΔΔC T ). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( A ) RT-qPCR for the expression of brnQ2 or brnQ3 was performed using cDNA from the wild-type parent, the brnQ1 null mutant, and the complementing strain after their incubation in a TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. ( B ) RT-qPCR for the expression of brnQ1 or brnQ3 was performed using cDNA from the wild type-parent, the brnQ2 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. ( C ) RT-qPCR for the expression of brnQ1 or brnQ2 was performed using cDNA from the wild-type parent, the brnQ3 null mutant, and the complementing strain after their incubation in TY medium for 2 h. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Expressing, Incubation, Bacteria, Centrifugation, Quantitative RT-PCR, Mutagenesis

    A comparison of PLC or PFO production levels for wild-type ATCC3624, its double null mutants, and the complementing strain when cultured in a TY medium. Western blot analyses of PLC ( A ) and PFO ( B ) levels in supernatants of the wild type, double null mutants, and their complementing strains in the early-stage culture (3 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right figure in panels A and B show the comparison of Western blot intensities determined by Image J. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of PLC or PFO production levels for wild-type ATCC3624, its double null mutants, and the complementing strain when cultured in a TY medium. Western blot analyses of PLC ( A ) and PFO ( B ) levels in supernatants of the wild type, double null mutants, and their complementing strains in the early-stage culture (3 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right figure in panels A and B show the comparison of Western blot intensities determined by Image J. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type.

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Cell Culture, Western Blot

    A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, or PLC and PFO production levels for ATCC3624 (WT) vs. the triple null mutant (TKO) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures incubated for up to 9 h at 37 °C. The results shown are representative of three repetitions. ( B ) The OD 600 of TY cultures after 24 h of incubation at 37 °C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio between 3 h samples (right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right graph in panel D shows the comparison of the Western blot band intensity ratio between 3 h samples using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PFO production differences among these strains in 7 or 24 h samples .

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: A comparison of post-inoculation changes in culture OD 600 , vegetative cell viability, or PLC and PFO production levels for ATCC3624 (WT) vs. the triple null mutant (TKO) when cultured in a TY medium. ( A ) Post-inoculation changes in the OD 600 of TY cultures incubated for up to 9 h at 37 °C. The results shown are representative of three repetitions. ( B ) The OD 600 of TY cultures after 24 h of incubation at 37 °C (top). The same samples were used for colony counting of viable vegetative cells (bottom). The mean values from three independent experiments are shown. The error bars indicate the S.D. ( C ) Western blot analyses of PLC levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. A comparison of the Western blot band intensity ratio between 3 h samples (right graph) using Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PLC production differences among these strains in 7 or 24 h samples . ( D ) Western blot analyses of PFO levels in the supernatants of panel A and B samples (3 h, 7 h and 24 h). The results shown are representative of three repetitions. The size of proteins in kiloDaltons (kDa) is shown on the left. The right graph in panel D shows the comparison of the Western blot band intensity ratio between 3 h samples using the Image J software. The mean values from three independent experiments are shown. The error bars indicate the S.D. * p < 0.05 relative to the wild type. Similar analyses did not detect any PFO production differences among these strains in 7 or 24 h samples .

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Comparison, Mutagenesis, Cell Culture, Incubation, Western Blot, Software

    C. perfringens strains used in this study.

    Journal: Toxins

    Article Title: BrnQ Branched-Chain Amino Acid Transporters Influence Toxin Production by, but Not Growth of, Clostridium perfringens Type A Strain ATCC3624

    doi: 10.3390/toxins17040187

    Figure Lengend Snippet: C. perfringens strains used in this study.

    Article Snippet: ATCC3624 , Wild type , Purchased from ATCC.

    Techniques: Mutagenesis

    Host range of phage Dolk21, lytic activity of PlyDolk21 and its enzymatic active domain PlyDolk21_EAD, and cell wall-binding activity of PlyDolk21_CBD.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Host range of phage Dolk21, lytic activity of PlyDolk21 and its enzymatic active domain PlyDolk21_EAD, and cell wall-binding activity of PlyDolk21_CBD.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Activity Assay

    Characterization of bacteriophage Dolk21. ( A ) TEM morphology of Dolk21 and ( B ) genome comparison among three C. perfringens phages Dolk21, CPAS-15, and CP3.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Characterization of bacteriophage Dolk21. ( A ) TEM morphology of Dolk21 and ( B ) genome comparison among three C. perfringens phages Dolk21, CPAS-15, and CP3.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Comparison

    Modular structures and expressions of PlyDolk21 from Dolk21. ( A ) Schematic representation of PlyDolk21, PlyDolk21_EAD, and PlyDolk21_CBD. ( B ) Amino acid sequence alignment of various C. perfringens phage endolysins: CP3 phage endolysin, Clo-PEP-1 phage endolysin, CPAS-15 phage endolysin, CPD4 phage endolysin. An asterisk (*) is marked every 10 amino acids. ( C ) SDS-PAGE analysis of purified PlyDolk21, PlyDolk21_EAD, and EGFP-fused PlyDolk21_CBD. M, standard molecular weight marker; PlyDolk21, purified PlyDolk21 fraction; PlyDolk21_EAD, purified PlyDolk21_EAD fraction; PlyDolk21_CBD, purified PlyDolk21_CBD fraction. ( D ) Lytic activities of PlyDolk21 and its EAD.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Modular structures and expressions of PlyDolk21 from Dolk21. ( A ) Schematic representation of PlyDolk21, PlyDolk21_EAD, and PlyDolk21_CBD. ( B ) Amino acid sequence alignment of various C. perfringens phage endolysins: CP3 phage endolysin, Clo-PEP-1 phage endolysin, CPAS-15 phage endolysin, CPD4 phage endolysin. An asterisk (*) is marked every 10 amino acids. ( C ) SDS-PAGE analysis of purified PlyDolk21, PlyDolk21_EAD, and EGFP-fused PlyDolk21_CBD. M, standard molecular weight marker; PlyDolk21, purified PlyDolk21 fraction; PlyDolk21_EAD, purified PlyDolk21_EAD fraction; PlyDolk21_CBD, purified PlyDolk21_CBD fraction. ( D ) Lytic activities of PlyDolk21 and its EAD.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Sequencing, SDS Page, Purification, Molecular Weight, Marker

    Stability of PlyDolk21 under various stress conditions was tested. Effects of ( A ) pH, ( B ) NaCl, and ( C ) temperature on the lytic activity of PlyDolk21 against C. perfringens ATCC 13124 cells. The means of triplicate experiments are represented in each column, and error bars indicate the standard deviation.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Stability of PlyDolk21 under various stress conditions was tested. Effects of ( A ) pH, ( B ) NaCl, and ( C ) temperature on the lytic activity of PlyDolk21 against C. perfringens ATCC 13124 cells. The means of triplicate experiments are represented in each column, and error bars indicate the standard deviation.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Activity Assay, Standard Deviation

    Binding activity of PlyDolk21_CBD to cell wall of C. perfringens in KH 2 PO 4 -NaOH buffer. Representative cell images show C. perfringens cells with PlyDolk21_CBD fused with EGFP. Panels from left to right show bright field, PlyDolk21_CBD with EGFP, and merged image.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Binding activity of PlyDolk21_CBD to cell wall of C. perfringens in KH 2 PO 4 -NaOH buffer. Representative cell images show C. perfringens cells with PlyDolk21_CBD fused with EGFP. Panels from left to right show bright field, PlyDolk21_CBD with EGFP, and merged image.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Binding Assay, Activity Assay

    Food applications of PlyDolk21 in ( A ) beef soup and ( B ) milk contaminated with C. perfringens ATCC 13124. Control, C. perfringens -contaminated food sample without PlyDolk21 (black); 0.1 μM, 0.5 μM, and 1.5 μM PlyDolk21-treated C. perfringens -contaminated beef soup or milk samples. PlyDolk21 was incubated with C. perfringens contaminated food samples for 4 or 24 h. Error bars present the standard deviations of three replicates. Asterisks indicate significant differences (*, p < 0.05; **, p < 0.01; ****, p < 0.0001).

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Food applications of PlyDolk21 in ( A ) beef soup and ( B ) milk contaminated with C. perfringens ATCC 13124. Control, C. perfringens -contaminated food sample without PlyDolk21 (black); 0.1 μM, 0.5 μM, and 1.5 μM PlyDolk21-treated C. perfringens -contaminated beef soup or milk samples. PlyDolk21 was incubated with C. perfringens contaminated food samples for 4 or 24 h. Error bars present the standard deviations of three replicates. Asterisks indicate significant differences (*, p < 0.05; **, p < 0.01; ****, p < 0.0001).

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: Control, Incubation

    Food applications of PlyDolk21_CBD in ( A ) KH 2 PO 4 -NaOH buffer, ( B ) beef soup, and ( C ) milk contaminated with C. perfringens ATCC 13124. Representative cell images show C. perfringens cells with PlyDolk21_CBD fused with EGFP. Top and bottom panels show bright field and PlyDolk21_CBD fused with EGFP, respectively.

    Journal: Antibiotics

    Article Title: Characterization of Clostridium perfringens Phage Endolysin PlyDolk21

    doi: 10.3390/antibiotics14010081

    Figure Lengend Snippet: Food applications of PlyDolk21_CBD in ( A ) KH 2 PO 4 -NaOH buffer, ( B ) beef soup, and ( C ) milk contaminated with C. perfringens ATCC 13124. Representative cell images show C. perfringens cells with PlyDolk21_CBD fused with EGFP. Top and bottom panels show bright field and PlyDolk21_CBD fused with EGFP, respectively.

    Article Snippet: C. perfringens , ATCC 3624 , − , +++ , + , +.

    Techniques: