[Construction of the prokaryotic expression plasmid pYNenv encoding the gp120 env gene of human immunodeficiency virus type 1 of yunnan strain].
The recombinant envelope protein of human immunodeficiency virus type 1(HIV-1) of Chinese Yunan epidemic strain was expressed in E. coli for confirming the antigenic domains of the protein. The recombinant prokaryotic expressive plasmid pYNenv was constructed by inserting the env DNA fragment which encode the gp120 of HIV-1 Yunnan strain into pBV220. The env DNA fragment was obtained by nested polymerase chain reaction with the HIV-1 infected patients peripheral blood macrophage and monocyte genomes from Yunan HIV-1 epidemic area as template. The recombinant plasmid pYNenv was confirmed by restriction enzyme analysis. The recombinant protein was expressed in E. coli DH10b by pYNenv after vibration culture at 30 degrees C for 20 hours then at 42 degrees C for 5 hours and confirmed by SDS-PAGE electrophoresis. The protein can specifically react with the serum of HIV-1 infected patient from Yunan epidemic area by Western blot assay. The result showed that the recombinant protein can be used as antigen for detection of HIV-1 membrane glycoprotein antibody as well as for the further study of the role of envelope protein in pathology.
Wang
B
B
Laboratory of Molecular Virology, Medical College of Qingdao University, Shandong 266021.
Lu
X
X
Shao
Y
Y
D000328:Adult; D003001:Cloning, Molecular; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics*; D015699:HIV Envelope Protein gp120 / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics; D011387:Prokaryotic Cells / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D000939:Epitopes; D015699:HIV Envelope Protein gp120; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins