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cexe -mcherry fusion  (GenScript corporation)

 
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    GenScript corporation cexe -mcherry fusion
    Unique Aat amino acid sequences were detected using PSI-BLAST. These sequences were used to identify the strains that encoded them in the NCBI identical protein groups. As the aat genes were present on contigs of whole genome sequencing projects, it was not possible to assess if a strain encoded a gene on the chromosome or plasmid. Instead, contigs were used to identify the complete Aat system. This analysis does not include strains that might encode the aat genes or aap / <t>cexE</t> on a separate genomic element. However, a total of 827 complete Aat systems were identified in the same nucleotide accession. The positions of these genes were used to assess the organisation of the aat operon. From this assessment five different classes of aat operon organisation were defined. Figure 1—source data 1. aatA gene positions. Figure 1—source data 2. aatB gene positions. Figure 1—source data 3. aatC gene positions. Figure 1—source data 4. aatD gene positions. Figure 1—source data 5. aatP gene positions. Figure 1—source data 6. aap or cexE gene positions. Figure 1—source data 7. Calculated distance between genes. Figure 1—source data 8. Distances and designations.
    Cexe Mcherry Fusion, supplied by GenScript corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cexe -mcherry fusion/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    cexe -mcherry fusion - by Bioz Stars, 2026-05
    90/100 stars

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    1) Product Images from "Glycine acylation and trafficking of a new class of bacterial lipoprotein by a composite secretion system"

    Article Title: Glycine acylation and trafficking of a new class of bacterial lipoprotein by a composite secretion system

    Journal: eLife

    doi: 10.7554/eLife.63762

    Unique Aat amino acid sequences were detected using PSI-BLAST. These sequences were used to identify the strains that encoded them in the NCBI identical protein groups. As the aat genes were present on contigs of whole genome sequencing projects, it was not possible to assess if a strain encoded a gene on the chromosome or plasmid. Instead, contigs were used to identify the complete Aat system. This analysis does not include strains that might encode the aat genes or aap / cexE on a separate genomic element. However, a total of 827 complete Aat systems were identified in the same nucleotide accession. The positions of these genes were used to assess the organisation of the aat operon. From this assessment five different classes of aat operon organisation were defined. Figure 1—source data 1. aatA gene positions. Figure 1—source data 2. aatB gene positions. Figure 1—source data 3. aatC gene positions. Figure 1—source data 4. aatD gene positions. Figure 1—source data 5. aatP gene positions. Figure 1—source data 6. aap or cexE gene positions. Figure 1—source data 7. Calculated distance between genes. Figure 1—source data 8. Distances and designations.
    Figure Legend Snippet: Unique Aat amino acid sequences were detected using PSI-BLAST. These sequences were used to identify the strains that encoded them in the NCBI identical protein groups. As the aat genes were present on contigs of whole genome sequencing projects, it was not possible to assess if a strain encoded a gene on the chromosome or plasmid. Instead, contigs were used to identify the complete Aat system. This analysis does not include strains that might encode the aat genes or aap / cexE on a separate genomic element. However, a total of 827 complete Aat systems were identified in the same nucleotide accession. The positions of these genes were used to assess the organisation of the aat operon. From this assessment five different classes of aat operon organisation were defined. Figure 1—source data 1. aatA gene positions. Figure 1—source data 2. aatB gene positions. Figure 1—source data 3. aatC gene positions. Figure 1—source data 4. aatD gene positions. Figure 1—source data 5. aatP gene positions. Figure 1—source data 6. aap or cexE gene positions. Figure 1—source data 7. Calculated distance between genes. Figure 1—source data 8. Distances and designations.

    Techniques Used: Sequencing, Plasmid Preparation

    ( A ) Culture supernatant of ETEC H10407 and aat mutants harbouring pCfaD grown in lysogeny broth (LB) supplemented with L-arabinose. Cells were removed from the culture supernatant and remaining protein precipitated with trichloroacetic acid. Protein samples were separated by Tris-tricine SDS-PAGE and stained with Coomassie or transferred to nitrocellulose for western blotting with polyclonal antibodies against CexE. ( B ) Whole-cell lysates of the aat mutants and parental strain separated by Tris-tricine SDS-PAGE. ( C ) Whole-cell lysates of ETEC H10407 grown in LB with or without azide. ( D ) Whole-cell lysates of EAEC 042, aap , aatD, and aatD complemented strains grown in DMEM (Dulbecco's Modified Eagle Medium) high glucose. The positions of uCexE, mCexE, uAap, mAap, and preCexE are indicated as appropriate. CexE and Aap were detected by western blotting (α-CexE and α-Aap, respectively) and RNA polymerase (α-RNAP) was included as a loading control where appropriate. Figure 2—source data 1. Western blot and Coomassie stained gels of culture supernatant fractions. Figure 2—source data 2. Western blot of whole-cell lysates derived from wild-type and mutant strains. Figure 2—source data 3. Western blot of whole-cell lysates after growth in sodium azide. Figure 2—source data 4. Western blot of whole-cell lysates showing Aap modification in the presence and absence of aatD .
    Figure Legend Snippet: ( A ) Culture supernatant of ETEC H10407 and aat mutants harbouring pCfaD grown in lysogeny broth (LB) supplemented with L-arabinose. Cells were removed from the culture supernatant and remaining protein precipitated with trichloroacetic acid. Protein samples were separated by Tris-tricine SDS-PAGE and stained with Coomassie or transferred to nitrocellulose for western blotting with polyclonal antibodies against CexE. ( B ) Whole-cell lysates of the aat mutants and parental strain separated by Tris-tricine SDS-PAGE. ( C ) Whole-cell lysates of ETEC H10407 grown in LB with or without azide. ( D ) Whole-cell lysates of EAEC 042, aap , aatD, and aatD complemented strains grown in DMEM (Dulbecco's Modified Eagle Medium) high glucose. The positions of uCexE, mCexE, uAap, mAap, and preCexE are indicated as appropriate. CexE and Aap were detected by western blotting (α-CexE and α-Aap, respectively) and RNA polymerase (α-RNAP) was included as a loading control where appropriate. Figure 2—source data 1. Western blot and Coomassie stained gels of culture supernatant fractions. Figure 2—source data 2. Western blot of whole-cell lysates derived from wild-type and mutant strains. Figure 2—source data 3. Western blot of whole-cell lysates after growth in sodium azide. Figure 2—source data 4. Western blot of whole-cell lysates showing Aap modification in the presence and absence of aatD .

    Techniques Used: SDS Page, Staining, Western Blot, Modification, Control, Derivative Assay, Mutagenesis

    His-tagged CexE was isolated from cexE or cexE aatD double mutants harbouring pACYC- cexE -6His grown in the presence or absence of 17-ODYA and separated by SDS-PAGE. An azide linked Alexa Fluor 488 was conjugated to the alkyne moiety present in 17-ODYA by CuAAC. The incorporation of 17-ODYA into the target protein was detected by fluorescence (green bands) and the image was overlaid on the image of the SDS-PAGE gel. A known lipoprotein YraP was used as a positive control.
    Figure Legend Snippet: His-tagged CexE was isolated from cexE or cexE aatD double mutants harbouring pACYC- cexE -6His grown in the presence or absence of 17-ODYA and separated by SDS-PAGE. An azide linked Alexa Fluor 488 was conjugated to the alkyne moiety present in 17-ODYA by CuAAC. The incorporation of 17-ODYA into the target protein was detected by fluorescence (green bands) and the image was overlaid on the image of the SDS-PAGE gel. A known lipoprotein YraP was used as a positive control.

    Techniques Used: Isolation, SDS Page, Fluorescence, Positive Control

    EAEC 042 and ETEC H10407 were grown in DMEM-high glucose (HG) or IMDM, respectively. After 2 hr of growth, 17-ODYA or an equal volume of DMSO was added. Cultures were grown for a further 3 hr. Triton X-100 was added to each culture to a final concentration of 0.1% for a final hour of growth. The culture supernatant was separated by centrifugation and filtered prior to TCA precipitation. Precipitated proteins were resuspended in 2% SDS. The incorporation of 17-ODYA into Aap and CexE was detected by CuAAC.
    Figure Legend Snippet: EAEC 042 and ETEC H10407 were grown in DMEM-high glucose (HG) or IMDM, respectively. After 2 hr of growth, 17-ODYA or an equal volume of DMSO was added. Cultures were grown for a further 3 hr. Triton X-100 was added to each culture to a final concentration of 0.1% for a final hour of growth. The culture supernatant was separated by centrifugation and filtered prior to TCA precipitation. Precipitated proteins were resuspended in 2% SDS. The incorporation of 17-ODYA into Aap and CexE was detected by CuAAC.

    Techniques Used: Concentration Assay, Centrifugation, TCA Precipitation

    The membrane fraction of ETEC H10407 and aat mutant harbouring pCfaD were separated by SDS-PAGE. Transferred to nitrocellulose and probed for the cytoplasmic protein RNAP, lipoprotein NlpE, and CexE using protein-specific antibodies.
    Figure Legend Snippet: The membrane fraction of ETEC H10407 and aat mutant harbouring pCfaD were separated by SDS-PAGE. Transferred to nitrocellulose and probed for the cytoplasmic protein RNAP, lipoprotein NlpE, and CexE using protein-specific antibodies.

    Techniques Used: Membrane, Mutagenesis, SDS Page

    E. coli BL21(DE3) was transformed with pET26b, or pET26b- cexE and pACYCDuet-1 or pACYC- aatD . Cultures were grown in lysogeny broth (LB) and protein production was stimulated with isopropyl β-D-1-thiogalactopyranoside (IPTG). Whole-cell lysate samples were taken and separated by Tris-tricine SDS-PAGE. CexE was detected using a polyclonal antibody. RNAP was used as a loading control. The position of mCexE and uCexE is indicated.
    Figure Legend Snippet: E. coli BL21(DE3) was transformed with pET26b, or pET26b- cexE and pACYCDuet-1 or pACYC- aatD . Cultures were grown in lysogeny broth (LB) and protein production was stimulated with isopropyl β-D-1-thiogalactopyranoside (IPTG). Whole-cell lysate samples were taken and separated by Tris-tricine SDS-PAGE. CexE was detected using a polyclonal antibody. RNAP was used as a loading control. The position of mCexE and uCexE is indicated.

    Techniques Used: Transformation Assay, SDS Page, Control

    ( A ) AatD from EAEC 042 and ETEC H10407 were aligned with Lnt from Escherichia coli MG1655. AatD sequences identified by PSI-BLAST were aligned and used to create a WebLogo. The catalytic residues of the carbon-nitrogen (C-N) hydrolase family are highlighted. ( B ) The structure of Lnt compared to the predicted structure of ETEC H10407 AatD. The catalytic residues of Lnt and AatD are highlighted in orange. ( C ) Magnified view of the catalytic site of Lnt with the predicted structure of AatD superimposed. Residues are numbered as they appear in Lnt. ( D ) WCL of E. coli BL21(DE3) expressing CexE in the presence of AatD and each of the four single site substitution derivatives of AatD. CexE was detected using an anti-CexE polyclonal antibody, AatD was detected with an anti-His tag antibody, and RNAP was used as a loading control.
    Figure Legend Snippet: ( A ) AatD from EAEC 042 and ETEC H10407 were aligned with Lnt from Escherichia coli MG1655. AatD sequences identified by PSI-BLAST were aligned and used to create a WebLogo. The catalytic residues of the carbon-nitrogen (C-N) hydrolase family are highlighted. ( B ) The structure of Lnt compared to the predicted structure of ETEC H10407 AatD. The catalytic residues of Lnt and AatD are highlighted in orange. ( C ) Magnified view of the catalytic site of Lnt with the predicted structure of AatD superimposed. Residues are numbered as they appear in Lnt. ( D ) WCL of E. coli BL21(DE3) expressing CexE in the presence of AatD and each of the four single site substitution derivatives of AatD. CexE was detected using an anti-CexE polyclonal antibody, AatD was detected with an anti-His tag antibody, and RNAP was used as a loading control.

    Techniques Used: Expressing, Control

    ( A ) SignalP result of CexE sequence from ETEC H10407 (E3PPH2_ECOH1) ( B ) WebLogo of the 5 N-terminal residues of Aap and CexE sequences post Sec signal sequence cleavage. ( C ) ETEC H10407 cexE mutants transformed with pACYC184 (Δ cexE ) or pACYC- cexE -6His with either the wild-type sequence (WT cexE ) or one of the first five amino acids mutated to glutamic acid. CexE was detected by polyclonal antibodies and RNAP was used as a loading control. The percentage of mCexE was determined from three biological replicates. Figure 7—source data 1. ETEC H10407 CexE sequence. Figure 7—source data 2. Aap and CexE sequences. Figure 7—source data 3. Aap and CexE sequences with signal sequences removed by SignalP. Figure 7—source data 4. T-coffee alignment of sequences. Figure 7—source data 5. Western blots of CexE production.
    Figure Legend Snippet: ( A ) SignalP result of CexE sequence from ETEC H10407 (E3PPH2_ECOH1) ( B ) WebLogo of the 5 N-terminal residues of Aap and CexE sequences post Sec signal sequence cleavage. ( C ) ETEC H10407 cexE mutants transformed with pACYC184 (Δ cexE ) or pACYC- cexE -6His with either the wild-type sequence (WT cexE ) or one of the first five amino acids mutated to glutamic acid. CexE was detected by polyclonal antibodies and RNAP was used as a loading control. The percentage of mCexE was determined from three biological replicates. Figure 7—source data 1. ETEC H10407 CexE sequence. Figure 7—source data 2. Aap and CexE sequences. Figure 7—source data 3. Aap and CexE sequences with signal sequences removed by SignalP. Figure 7—source data 4. T-coffee alignment of sequences. Figure 7—source data 5. Western blots of CexE production.

    Techniques Used: Sequencing, Transformation Assay, Control, Western Blot

    Aap and CexE sequences were aligned using ClustalO. The alignment was then placed into WebLogo.
    Figure Legend Snippet: Aap and CexE sequences were aligned using ClustalO. The alignment was then placed into WebLogo.

    Techniques Used:

    Enterotoxigenic Escherichia coli (ETEC) H10407Δ cexE pCfaD was transformed with pACYC184, pACYC- cexE , or pACYC- cexE G20E. A double mutant of cexE and aatD harbouring pCfaD was transformed with pACYC- cexE . Both WCL and culture supernatant samples of these strains grown in the presence of arabinose were taken. Supernatant proteins were isolated by TCA precipitation after removal of cells by centrifugation and filtration. Protein samples were separated by SDS-PAGE and transferred to nitrocellulose for western blotting for RNAP and CexE. Figure 7—figure supplement 2—source data 1. Coomassie stained gel and western blots of secreted CexE.
    Figure Legend Snippet: Enterotoxigenic Escherichia coli (ETEC) H10407Δ cexE pCfaD was transformed with pACYC184, pACYC- cexE , or pACYC- cexE G20E. A double mutant of cexE and aatD harbouring pCfaD was transformed with pACYC- cexE . Both WCL and culture supernatant samples of these strains grown in the presence of arabinose were taken. Supernatant proteins were isolated by TCA precipitation after removal of cells by centrifugation and filtration. Protein samples were separated by SDS-PAGE and transferred to nitrocellulose for western blotting for RNAP and CexE. Figure 7—figure supplement 2—source data 1. Coomassie stained gel and western blots of secreted CexE.

    Techniques Used: Transformation Assay, Mutagenesis, Isolation, TCA Precipitation, Centrifugation, Filtration, SDS Page, Western Blot, Staining

    ( A ) CexE-6His was isolated from a cexE mutant and a cexE aatD double mutant and subjected to LC-MS/MS. CexE and pro-CexE were trypsinated and separated by HPLC. ( B ) The indicated peak was subjected to MS/MS to identify the amino acid sequence. Figure 8—source data 1. HLPC of mCexE. Figure 8—source data 2. HLPC of uCexE. Figure 8—source data 3. Peptide results.
    Figure Legend Snippet: ( A ) CexE-6His was isolated from a cexE mutant and a cexE aatD double mutant and subjected to LC-MS/MS. CexE and pro-CexE were trypsinated and separated by HPLC. ( B ) The indicated peak was subjected to MS/MS to identify the amino acid sequence. Figure 8—source data 1. HLPC of mCexE. Figure 8—source data 2. HLPC of uCexE. Figure 8—source data 3. Peptide results.

    Techniques Used: Isolation, Mutagenesis, Liquid Chromatography with Mass Spectroscopy, Tandem Mass Spectroscopy, Sequencing

    Plasmid encoding mCherry with the CexE signal sequence followed by none, one (G), two (GG), or three (GGG) glycine residues post signal sequence was produced in an ETEC H10407 cexE mutant grown in the presence of 17-ODYA. The mCherry proteins were isolated using a C-terminal 8 His tag and azide linked Cy5 was incorporated into mCherry proteins using CuAAC. The acylation of mCherry was detected using fluorescence. Figure 9—source data 1. ODYA labelling of chimeric proteins.
    Figure Legend Snippet: Plasmid encoding mCherry with the CexE signal sequence followed by none, one (G), two (GG), or three (GGG) glycine residues post signal sequence was produced in an ETEC H10407 cexE mutant grown in the presence of 17-ODYA. The mCherry proteins were isolated using a C-terminal 8 His tag and azide linked Cy5 was incorporated into mCherry proteins using CuAAC. The acylation of mCherry was detected using fluorescence. Figure 9—source data 1. ODYA labelling of chimeric proteins.

    Techniques Used: Plasmid Preparation, Sequencing, Produced, Mutagenesis, Isolation, Fluorescence


    Figure Legend Snippet:

    Techniques Used: Recombinant, Plasmid Preparation, Expressing



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