Leishmania donovani amastin recombinant eukaryotic expression plasmid and expression of.docVIP

 PAGE \* MERGEFORMAT 16 Leishmania donovani amastin recombinant eukaryotic expression plasmid and expression of Authors: Li Jinfu, Chen Jianping, YANG Zhi-wei, Tian, Ying Ma, HU Xiao-su [Keywords:] Leishmania donovani;, non-flagellar protein gene;, in vitro expression Construction and expression of recombinant eucaryotic expression plasmid of amastin gene of Leishmania Donovani [Abstract] AIM: To construct recombinant eukaryotic expression plasmid of amastin gene of Leishmania Donovani anddetectexpressionofthegeneinNIH3T3cells.METHODS: Amastin gene was amplified from nuclear DNA of Leishmania Donovani isolates and cloned into an eukaryotic expression vector pcDNA31 (). The recombinant plasmid was named pcDNA31amastin. NIH3T3 cell was transfected by pcDNA31amastin. Transient and stable expression of amastin gene were detected by immunofluoresence and RTPCR. 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 pcDNA31amastin 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. [Keywords] Leishmania Donovani; amastin gene; express in vitro [Abstract] Objective: To construct Leishmania donovani amastin (amastin) gene encoding the eukaryotic expression recombinant plasmid pcDNA31amastin, and to study its expression in NIH3T3 cells. Methods: Extraction of Leishmania donovani genomic DNA for PCR amplification. Will be amplified amastin gene fragment into plasmid pcDNA31 (), the eukaryotic expression recombinant plasmid pcDNA31amastin. To pcDNA31amastin transfected NIH3T3 cells, using immunofluorescence staining and RTPCR were identified pcDNA31amastin transient expression and stable expression. Results: In the cell membrane and the cells were observed in strong green fluorescence, indicating successful pcDNA31amastin into NIH3T3 cells