TI:Screening and cloning of the genes of protein interacting with the N2terminal protein of hepatitis B virus DNA polymerase by yeast2two hybrid technique
AU:CHEN Guofeng, WANGLin , CHENGJun , ZHANGLingxia ,LI Li , ZHANG Jian , SHAO Qing , JI Dong.
AD:The 4 th Department , The No1302 Hospital of The People’s Liberation Army , Beijing 100039 ,China Corresponding author : CHENG Jun , Tel : 010266933391 , Fax : 010263801283 , E2mail : cj @ genetherapy1com1cn
【Abstract】　Objective 　To screen and clone the genes of protein interacting with the N2terminal protein (TP) of hepatitis B virus DNA polymerase1 Methods 　TP was amplified by polymerase chain reaction(PCR) and TP bait plasmid was constructed by using yeast two2hybrid system 3 , then transformed into yeast AH 1091 The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2×YPDA medium1 Diploid yeast was plated on synthetic dropout medium(SDP2Trp2Leu2His2Ade) and that containing X2α2GAL for selecting two times and screening1 Plasmids were extracted from blue colonies , and sequence analysis was performed by bioinformatics1 Results 　Forty2seven clones were obtained , these clones included human P36956 sterol regulatory element binding protein21 , RNA polymerase Ⅱ subunit hsRPB7 mRNA , asialoglycoprotein receptor 2 , transcript variant 3 , ceruloplasmin (ferroxidase) , transmembrane 4 superfamily member 2 and 19 of the hypothetical proteins and so on1 Conclusion 　Genes encoding TP interacting proteins in hepatocytes were successfully cloned and the results suggest that TP has a wide variety of biological functions1
【Key words】　Hepatitis B virus ; DNA; Yeasts