[Screening and cloning gene of a hepatocyte protein interacting with HCV NS5ATP4A protein].
OBJECTIVE
To screen and clone hepatocyte protein interacting with hepatitis C virus NS5ATP4A protein for studying its biological functions.


METHODS
Bait plasmids of hepatitis C virus NS5ATP4A were constructed. After verifying that hepatitis C virus NS5ATP4A protein could be steadily expressed in AH109 yeast strain, yeast-two hybrid assay was performed by mating AH109 with Y187 which pre-transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X-a-gal activity. Nineteen yeast colonies which grew on QDO and had a-gal activity were obtained, and then the library plasmids were extracted and sequenced.


RESULTS
Seven genes were screened out and one of them was a formerly unknown gene. They were associated with RNA synthesis, protein translation, cell cycling and tumor immunity.


CONCLUSIONS
NS5ATP4A binding proteins were successfully screened, which offers new clues for further studying the signal transduction pathway of NS5ATP4A and the pathogenic mechanism of HCV.
Zhang
Lian-feng
LF
Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. lfzhang1999@126.com
Cheng
Jun
J
Li
Ji-chang
JC
Chen
Li-dong
LD
Liu
Wei
W
D001483:Base Sequence; D003001:Cloning, Molecular; D015723:Gene Library; D016679:Genome, Viral; D016174:Hepacivirus / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D017385:Sequence Homology; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase