Healthy APOBEC3G gene and the eukaryotic expression vector and identification of.docVIP

 PAGE \* MERGEFORMAT 13 Healthy APOBEC3G gene and the eukaryotic expression vector and identification of Of: Gan Yimin, Fan Yun, Shi Zhen, Tang Ren Xian, Cheng Kui Yang [Abstract] Objective To clone the human apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G (apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3G, APOBEC3G) gene cDNA, APOBEC3G gene construct eukaryotic expression vector and in vitro expression and identification. Methods RT-PCR technique from human peripheral blood mononuclear cells cloned APOBEC3G gene, the gene was inserted into pcDNA3.1 eukaryotic expression vector pcDNA3.1-A3G construct eukaryotic expression plasmid and the recombinant plasmid by liposome pcDNA3.1-A3G transfected into HepG2 and HL7702 cells by immunocytochemistry, Western blot used to detect the expression of APOBEC3G eukaryotic. Results restriction enzyme digestion and sequencing results showed that, APOBEC3G was successfully constructed the eukaryotic expression vector. immunocytochemistry and Western blot results were transfected with pcDNA3.1-A3G in HepG2 and HL7702 cells with high expression of APOBEC3G protein were transfected with empty vector pcDNA3.1 and untransfected HepG2 and HL7702 cells with low expression of APOBEC3G protein or expression. Conclusion Construction of the human apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G eukaryotic expression vector pcDNA3.1-A3G, APOBEC3G for the further study of the role of anti-HBV experimental basis. [Keywords:] APOBEC3G; cloning; eukaryotic expression vector Abstract: Objective To clone the cDNA of human apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3G (APOBEC3G) gene and construct the eukaryotic expression vector for the in vitro expression and identification. Methods The coding region of APOBEC3G gene was amplified by RT- PCR from peripheral blood mononuclear cells (PBMCs) of healthy humans, and then the gene fragment was inserted into the recom