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Cloning and sequence analysis of an amastin coding gene from Leishmania major Abdou
1Gene Therapy Research Center, Institute of Infectious Diseases, 302 Hospital of PLA, Beijing 100039, China.
Objective:
To clone a surface protein encoding gene from Leishmania major Abdou parasites.
Methods:
Using Trypanosotidae cruzi (T. cruzi) amastin DNA sequence as a reference, computer search was done on Genbank and dbEST data bases with BLASTN path. A Leishmania major Abdou DNA library has been established and screened 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. Leishmania major Abdou DNA library was screened using specific probes synthesized according to 309 nt DNA sequence of T. cruzi amastin gene, and full-length coding sequence for Leishmania major Abdou 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 Abdou and T. cruzi amastins.
Conclusion:
We have cloned the full length amastin coding DNA for Leishmania major Abdou.