Cloning and Sequencing of Leishmania major Thiol-Specific- Antioxidant Antigen (TSA) Gene

Background: Leishmaniasis is caused by parasitic protozoa of the genus Leishmania which, in the infected host are obligate intracellular parasite. TSA is the immuno-dominant antigen of Leishmania major which is considered as the most promising molecule for a recombinant or DNA vaccine against leishmaniasis. Methods: Genomic DNA of TSA protein was extracted and amplificated as a template. Then the PCR product was cloned into pTZ57R/T vector. Finally, the recombinant plasmid was extracted from transformed Escherichia coli (TG1 strain) and sequenced. Results: MRHO/IR/75/ER (an Iranian strain) of L. major and TSA gene (Accession number LmjF15.1080) were used. Sequence analysis of cloned TSA gene into pTZ57R/T vector showed high homology of 90% with LmjF15.1080 (TSA gene) and strain LV39 (Accession no. AF069386) and strain Friedlin (Accession no.AF044679). Conclusion: We cloned TSA gene of L. major successfully. Recombinant plasmid was confirmed. It is ready to express recombinant protein for further studies.