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[Cloning and sequence analysis of a gene encoding amastin from Leishmania major]
1Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039.
Objective:
To clone a gene encoding surface protein from Leishmania major.
Methods:
Using T. cruzi amastin DNA sequence as a reference, computer search was done on GenBank and dbEST databases by using BLAST path. A Leishmania major DNA library has been constructed and screened by in situ colony hybridization.
Results:
A 309nt DNA fragment from Leishmania major was found in dbEST. Leishmania major DNA library was screened using specific primers synthesized according to 309 nt DNA sequence, and a full-length coding sequence for Leishmania major amastin was cloned. The coding sequence consisted of 552 nt, and translated into 183 amino acid residues. The homology is 23.5% at amino acid sequence level between Leishmania major and T. cruzi amastins.
Conclusion:
A full length amastin coding gene for Leishmania major has been cloned.