Cloning and sequence analysis of an amastin coding gene from Leishmania major Abdou

Objective To clone a surface protein encoding gene from Leishmania major Ab dou parasites. Methods Using Trypanosotidae cruzi (T. cruzi) amastin DNA sequence as a ref erence, computer search was done on Genbank and dbEST data bases with BLAST N path. A Leishmania major Abdou DNA library has been established and scree ned by in situ colony hybridization. Results No homology sequence to T, cruzi amastin was found in Genbank, but a 309 nt DNA fragment from Leishmania major (L. major) existed in bEST. Lei shmania major Abdou DNA library was screened using specific probes synthesi zed according to 309 nt DNA sequence of T. cruzi amastin gene, and full-len gth coding sequence for Leishmania major Abdou amastin was cloned. The codi ng sequence consisted of 552 nt, and translated into 183 amino acid residue s. The homology is 23.5% at amino acid sequence level between Leishmania ma jor Abdou and T. cruzi amastins. Conclusion We have cloned the full length amastin coding DNA for Leishmania major Abdou.