Cloning and sequencing of Leishmania major thiol-specific-antioxidant antigen [TSA] gene

ABSTRACT

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. 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. 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]. We cloned TSA gene of L. major successfully. Recombinant plasmid was confirmed. It is ready to express recombinant protein for further studies