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Cloning and expression of LACK gene of Leishmania donovani


Cloning and expression of LACK gene of Leishmania donovani



Chinese Journal of Zoonoses 18(5): 42-44



Aims To clone the LACK gene of Leishmania donovani and express it in Escherichia coli ( E. coli ). Methods DNA fragment encoding the LACK gene of Leishmania dmovani was obtained from the recombinant plasmid T-LACK by PCR.TI gene was cloned into expressed plasmid pQESI, and then was transformed into E. coli MI5 for expression and purification. The expressed protein was analyzed by SDS - PAGE followed by Western blotting and immuno-staninig. Results 1. We obtained the protein by three methods from E .coli MI5 transformed with the recombinant plasmid PQE31-LACK.The specific protein band with molecular weight of 36kDa was detected on SDS-PAGE both in the purified protein and solution of inclusion body except the who protein of E. coli. 2. The purified protein was identified by immuno-staining. Conclusion Efficient expression of LACK purified pn protein ol c. coil. 2. I he punlied protein was identifaed by immuno-staining. Conclusion Efficient expression of LACK purified protein has been achieved in E. coli and the protein has immunocoinpetence.

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