Aim: To construct recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani and detect expression of the gene in NIH3T3 cells.
Methods: Amastin gene was amplified from nuclear DNA of Leishmania Donovani isolates and cloned into an eukaryotic expression vector pcDNA3.1(+). The recombinant plasmid was named pcDNA3.1-amastin. NIH3T3 cell was transfected by pcDNA3.1-amastin. Transient and stable expression of amastin gene were detected by immunofluoresence and RT-PCR.
Results: It was found that there was high green fluorescence on the cell membrane and inside the cell. It showed that NIH3T3 cell was transfected by pcDNA3.1-amastin successfully.
Conclusion: A recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani was successfully constructed, and can be expressed stably in the NIH3T3 cells.