The study on genes trans-regulated by hepatitis C virus core protein-binding protein 6

Objective: To analyze the expression profile of genes trans-regulated by HCV core protein-binding protein 6 （HCBP6） gene.Methods: Human HCBP6 full-length encoding gene and its amino acid sequences was identified using bioinformatics method and the recombined expression plasmid pcDNA3.1（-）-HCBP6 was constructed. Suppression subtractive hybridization （SSH） technique was employed to detect the mRNA from HepG2 cells transfected with pcDNA3.1（-）-HCBP6 and the empty vector, respectively. The twice enriched PCR products were subcloned into T/A vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results: According to yeast two-hybrid screening and the bioinformatics analysis results, human HCBP6 cDNA sequence was identified. The encoding sequence for human HCBP6 consists of 456 nt and its protein consists of 152 aa. The amplified subtractive library contains 30 positive clones. Colony PCR analysis shows there are 24 clones containing 200-1 000 bp inserts. Sequence analysis was performed and 11 kinds of encoding sequences were achieved. Conclusion: The bioinformatics combined yeast two hybrid methods is a powerful method for the screening and analysis of genes of hepatocyte protein interacting with HCV core protein. The findings obtained using SSH provide significant data for preliminary understanding the biological function of a new identified gene.