1.Screening and cloning gene of hepatocyte protein interacting with hepatitis C virus core protein.
Ke LI ; Lin WANG ; Jun CHENG ; Lingxia ZHANG ; Huijuan DUAN ; Yinying LU ; Jizhen YANG ; Yan LIU ; Xiaobing XIA ; Gang WANG ; Jing DONG ; Li LI ; Yanwei ZHONG ; Yuan HONG ; Jumei CHEN
Chinese Journal of Experimental and Clinical Virology 2002;16(4):351-353
OBJECTIVETo clone the unknown gene of hepatocyte protein interacting with hepatitis C virus core protein.
METHODSUsing the yeast dual hybrid system 3, bait plasmids of hepatitis C virus core were constructed. After identifying hepatitis C virus core protein that could stably expressed in AH109 yeast strains, we performed yeast two hybrid by mating AH109 with Y187 that transformed with liver cDNA library plasmids pACT2 and then plated on quadrople dropout (QDO) medium and assayed for alpha-gal activity. The genes of yeast colonies that could grow on QDO and had alpha-gal activity were sequenced.
RESULTSAmong the 30 positive colonies, we blasted the gene of the sixth colony; we coined human hepatitis C virus binding protein 6(Hu Hcbp6) with Genbank, realized that the Hu Hcbp6 shares as much as 98% homology with two cDNA without knowing functions. We have proved that Hu Hcbp6 could interact with hepatitis C virus core protein.
CONCLUSIONSHepatitis C virus core binding protein (Hu Hcbp 6 Genbank number: AY032594) was successfully cloned and identified. The study partly paved the way for investigating physiological function of the Hu Hcbp6.
Cloning, Molecular ; DNA, Complementary ; genetics ; DNA-Binding Proteins ; genetics ; Hepacivirus ; Humans ; Molecular Sequence Data ; Plasmids ; Sequence Analysis, DNA ; Transfection ; Two-Hybrid System Techniques ; Viral Core Proteins ; genetics ; metabolism ; Yeasts ; genetics
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