| Hepatitis B virus (HBV) DNA Polymerase (HBV Pol) is the longest ORF in HBV and overlaps with the S, C, X genes. The P protein coded by HBV Pol has 3 functional domains and 1 spacer (SP) that are lined up for terminal protein (TP), SP, reverse transcriptase/polymerase (RT/PR) and RNase H. Up to now, the HBV Pol and its functional domains have not been clearly clarified. In order to investigate the effects of viral protein on hepatocytes, the genes of protein interacted with proteins of functional domain proteins of HBV Pol were screened and cloned using yeast two-hybrid system. Meanwhile, microarray assay was applied to study the difference in gene expression in human hepatoblastoma cell line HepG2 cells transfected with functional domain proteins of HBV Pol expressing plasmid.I : Sequence specific primers were designed and synthesized and the TP, PR and RNase H coding DNA fragment was amplified, respectively, with polymerase chain reaction (PCR) technique using G318A7 AF384372 plasmid containing the full length of HBV Pol cDNA as the template. Using MATCHMAKER Two-Hybrid System 3, the bait plasmids of TP, PR and RNase H were constructed by ligating these genes into pGBKT7, then the vectors were transformed into yeast AH109 by the lithium acetate (LiAc)-mediated method. The proteins were expressed and confirmed by Western blotting. The transformed yeast AH109/pGBKT7-TP was mated with yeast Y187 containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade, QDO and SD/-Trp-Leu-His, TDO) and QDO containing X- α -gal for selecting two times and screening. Yeast plasmids were extracted from blue colonies, and sequence analysis was performed by bioinformatics methods.A total of 47 clones were obtained from TP study. They included Human P36956 sterol regulatory element binding protein-1 (SREBP1), RNA polymerase II subunit hsRPB7 mRNA, Ceruloplasmin (ferroxidase) (CP) , alcohol dehydrogenase class I Y...
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