[Screening and cloning gene of hepatocyte protein interacting with hepatitis C virus core protein].
OBJECTIVE
To clone the unknown gene of hepatocyte protein interacting with hepatitis C virus core protein.


METHODS
Using 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.


RESULTS
Among 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.


CONCLUSIONS
Hepatitis 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.
Li
Ke
K
Gene Theraphy Research Center,Institute of Infectious Diseases, The 302nd Hospital of PLA, Beijing 100039, China.
Wang
Lin
L
Cheng
Jun
J
Zhang
Lingxia
L
Duan
Huijuan
H
Lu
Yinying
Y
Yang
Jizhen
J
Liu
Yan
Y
Xia
Xiaobing
X
Wang
Gang
G
Dong
Jing
J
Li
Li
L
Zhong
Yanwei
Y
Hong
Yuan
Y
Chen
Jumei
J
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D016174:Hepacivirus; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D017422:Sequence Analysis, DNA; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D015003:Yeasts / Q000235:genetics
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D014758:Viral Core Proteins