l tarentolae parrot tar  (ATCC)

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: CRISPR/Cas9 tagging and expression of LtVtc1 and LtVtc4. (A) Schematic representation of LtVtc1 and LtVtc4. SPX: SYG1/Pho81/XPR domain, CD: Catalytic domain, TMH: Transmembrane helix domain. (B) PCR verification of mNG-tagged LtVtc1 and LtVtc4 showing a band at ∼2200 bp, absent in WT. (C) Fluorescent detection of mNG-LtLtVtc1, and LtVtc4 expressed in L. tarentolae ; Coomassie blue staining of WT L. tarentolae , mNG-LtVtc1, mNG-LtVtc4 confirms comparable total protein levels, ensuring consistent sample loading for fluorescent detection. (D) Fluorescence microscopy micrographs of mNG-LtLtVtc1 and mNG-LtVtc4 with mPPase in L.tarentolae (Pearson’s r = 0.85 and 0.83, respectively), scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: CRISPR, Expressing, Staining, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged by immunofluorescence microscopy. mNG-LtVtc4 colocalises with mCh-LtVtc1 (Pearson’s r = 0.83) as well as with mCh-LtPho91, mCh-LtCa²⁺-ATPase, mCh-LtV-H⁺-ATPase_A, and mCh-LtZnT, showing Pearson’s correlation coefficients of 0.87, 0.83, 0.87, and 0.84, respectively. A lower degree of colocalisation is observed with mCh-IP₃R (r = 0.67). Scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Expressing, Immunofluorescence, Microscopy

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet:

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Colocalisation analysis of L. tarentolae expressing mNG-LtVtc4 and mCh-tagged LtVBP1, LtVBP2, and LtVBP3 by immunofluorescence microscopy. All three proteins show colocalisation with LtVtc4 in acidocalcisomes. Pearson’s correlation coefficients with LtVtc4 are 0.83 for LtVBP1, 0.85 for LtVBP2, and 0.82 for LtVBP3. Scale bar: 5 µm.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Expressing, Immunofluorescence, Microscopy

Journal: bioRxiv

Article Title: Novel Binding Partners of the Vacuolar Transporter Chaperone (VTC) complex in Acidocalcisomes of Leishmania tarentolae

doi: 10.1101/2025.09.23.677757

Figure Lengend Snippet: Pulldown assay of novel VTC binding partners with mNG-LtVtc4 in L. tarentolae. (A-C) SDS-PAGE with fluorescence detection shows capture of mCh-LtVBP1, mCh-LtVBP2, and mCh-LtVBP3 by mNG-LtVtc4 using mNeonGreen-Trap agarose beads, as indicated by signal in the bound fraction. (D) Pulldown of mNG-LtVtc4 with mCh-LtVBP3 using ChromoTek RFP-Trap beads confirms interaction. Lane labels: I – input; FT – flow-through; W3 – third wash; B – bound fraction. Green bands correspond to mNG-LtVtc4; some I and B lines appear as yellowish due to partial detection of the mNG tag in the 532 nm channel ( S6 Fig ). Red bands correspond to mCherry-tagged proteins. The fluorescence signal marked by a white asterisk in the gel represents free mCh tag, visible only in the input due to partial degradation.

Article Snippet: GO analysis of biological processes, cell components, and molecular functions was performed on the 412 overlapping LtVtc1 and LtVtc4 interactors using the TriTrypDB GO analysis tool [ ] with the complete L. tarentolae parrot tar II 2019 (ATCC 30267) genome as the reference.

Techniques: Binding Assay, SDS Page, Fluorescence

Journal: Iranian Biomedical Journal

Article Title: Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum

doi: 10.52547/ibj.25.5.349

Figure Lengend Snippet: Growth curves of the L. tarentolae -PpSP15-EGFP parasite cultured in two different media with 1× 10 7 /ml inoculation in 25-cm 2 flasks at 26 °C for a five-day period. (A) Growth rate evaluation of L. tarentolae -PpSP15-EGFP parasites (parasite count/ml) in CSFM medium compared to M199 5% FBS. (B) Growth rate evaluation of L. tarentolae -PpSP15-EGFP parasites (optical density at 600 nm/ml) in CSFM and M199 5% FBS. The experiment was repeated three times, and values are pooled in the Figure. Bars show the mean ± SD. The significant difference in parasite growth rate for different media was determined by t -test analysis and demonstrated as asterisk at the indicated time points ( p < 0.05 denoted as * ). At the time points that are not marked with the asterisk, no significant difference was observed ( p > 0.05)

Article Snippet: Parasite culture Three types of L. tarentolae , including the wild-type L. tarentolae Tar II (ATCC 30.267) strain, EGFP-expressing L. tarentolae ( L. tarentolae -EGFP) [ ] , and recombinant L. tarentolae -PpSP15-EGFP [ ] (as previously available), as well as wild-type L. major strain (MRHO/IR/75/ER) were used.

Techniques: Cell Culture

Journal: Iranian Biomedical Journal

Article Title: Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum

doi: 10.52547/ibj.25.5.349

Figure Lengend Snippet: Determination of EGFP expression in CSMF medium in comparison with M199 supplemented with 5% FBS. (A) Assessment of EGFP expression by epifluorescence microscopy, indicating the EGFP expression of L. tarentolae -EGFP and L. tarentolae -PpSP15-EGFP promastigotes in the logarithmic and stationary phases in both CSFM and M199 5% FBS media. (b) Flow cytometry analysis of recombinant L. tarentolae -PpSP15-EGFP (right panel), L. tarentolae EGFP as a positive control (middle panel), and the L. tarentolae wild type as a negative control (left panel) using a FITC detector in CSFM and M199 5% FBS at logarithmic (upper panel) and stationary (lower panel) phases. (C) Western blot analysis to confirm the expression of PpSP15-EGFP by recombinant L. tarentolae -PpSP15-EGFP parasite in CSFM and M199 with 5% FBS using an anti-GFP antibody. A 42-kDa band relating to the expression of PpSP15-EGFP protein in concentrated supernatant of L. tarentolae -PpSP15-EGFP parasite was detected in M199 with 5% FBS (lanes 2 and 3) and CSFM medium (lanes 5 and 6), respectively at logarithmic and stationary phases. A 27-kDa band indicating EGFP protein in the L. tarentolae -EGFP parasite (lane 1) and wild type form of L. tarentolae parasite as negative control was cultivated in M199 with 5% FBS (lane 4)

Article Snippet: Parasite culture Three types of L. tarentolae , including the wild-type L. tarentolae Tar II (ATCC 30.267) strain, EGFP-expressing L. tarentolae ( L. tarentolae -EGFP) [ ] , and recombinant L. tarentolae -PpSP15-EGFP [ ] (as previously available), as well as wild-type L. major strain (MRHO/IR/75/ER) were used.

Techniques: Expressing, Comparison, Epifluorescence Microscopy, Flow Cytometry, Recombinant, Positive Control, Negative Control, Western Blot

Journal: Iranian Journal of Basic Medical Sciences

Article Title: In silico analysis and expression of a new chimeric antigen as a vaccine candidate against cutaneous leishmaniasis

doi: 10.22038/ijbms.2020.45394.10561

Figure Lengend Snippet: SDS-PAGE analysis of the level of expression of chimeric sequence that successfully subcloned to pLEXY-neo2 and expressed in Lishmania tarentolae . Lane 1, Protein molecular weight marker (10–140 kDa); Lanes 2: Logarithmic phase secretory sample, Lanes 3: Stationary phase secretory sample, Lanes 4: Logarithmic phase cytosolic sample, Lanes 5: Stationary phase cytosolic sample and Lanes 5: Control ( L. tarentolae secretory sample) (A). Western blot analysis ( B)

Article Snippet: Transfection of pLEXY-TLGL into L. tarentolae The L. tarentolae Tar II (ATCC 30143) strain was grown in RPMI-1640 medium (Gibco) treated with 10% fetal calf serum (FCS, Gibco) affected by heat inactivation (pH: 7.2 and 26 o C).

Techniques: SDS Page, Expressing, Sequencing, Molecular Weight, Marker, Control, Western Blot