A Study On Proteins Binding With HCV NS2TP Promoter

Hepatitis c virus is the important pathogen for chronic liver disease which includes chronic active hepatitis, liver cirrhosis and hepatocellular carcinoma. Recently, the clinical observations find that, compared with other viral hepatitis, hepatitis C develops more serious chronic liver disease. HCV RNA translates one polypeptide which cleavage into ten proteins, that is, core, El, E2, P7, NS2, NS3, NS4a, NS4b, NS5a, NS5b. At present, some studies indicate the ORF of HCV core could move to a novel one which is named F protein with 17 KD. All of these proteins play differently important roles in HCV replication and propagation, but some specific functions are not clear.HCV NS2 could up-regulate and down-regulate the expression of many kinds of liver cell Cytokines, that is important in the happeness and development of carcinoma and could be one of the mechanism of hepatitis and hepatocellular carcinoma .NS2TP is a novel NS2 transregulated protein in this study,it can combine with CYP2E1, Decorin, P68, beta-2-microglobulin, CPN2, metallothionein 2A, insulin-like growth factor binding protein 4 and FGG et al.these proteins are important cytokines in the regulation of immunity,so, NS2TP is a significant immune protein. And it is a protein of secreting type,which make it easy binding with drugs and being drugs.By using HCV NS2TP promoter DNA as the selective molecule, the T7 select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment and cloned into pGEM-Teasy vector. After sequencing and homology searching, proteins binding with HCV NS2TP promoter was definited.Sequence analysis was performed in tirty positive plaque, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether twelve coding sequences were gotten. NS2TP promoter binding proteins were screened. These proteins need to be further studied and will be usefully for further study of the expression and regulation mechanism of NS2.