The Preliminary Study Of The Regulation Mechanism Of HCV Core Protein Binding Protein 6 (HCBP6) At Transcriptional Level

HCV is one of main reason of the chronic hepatitis, hepatic cirrhosis and carcinomatous, almost 250,000-350,000 succumb to the HCV and related disease.So,it is no denied HCV is a serious problem of socity and public health, nevertheless, we have no moer effcctive therapeutic metheod except the united of interferon and ribovirin, the worse matter is that we have no vaccine for HCV. One of the main reason of the lag of the revent and therapy is that we can not relize the molecular biology mechanism of HCV in a more deep lever, so the research of the molecular biology mechanism of HCV is the hot spot of this area.HCBP6 is a protein that can conioin with the HCVcore, which we screened from the liver cDNA Lib, we find that HCBP6 can up-regulation and down regulation the expression of some gene, and this differential expression be correlated with the signal transduction, hyperplasia, differentiation and the growth control of the cell. So it is necessary for us to research the HCBP6 to move forward a single step.The regulation in the transcriptional level is more impotant to the HCV, because hepatitis C virus is an RNA virus,it different with the pathogenic mechanism of the HBV,can not to integrate with the geneome of hepatic cell,because it will not existence the stage of DNA.the chief pathogenic mechanism are the combination of viral protein and the hepatic cell protein,and the trans-regulation by this combination.Objective My research is to study the regulation mechanism of HCV core protein binding protein 6 (HCBP6), which a new gene screened by our research team at earlier experiments, at transcriptional level.Methods I use the luciferase report system as a technology platform. The specific process is as follows. Firstly, I determine the HCBP6 promoter by bioinformatics. Secondly, I identificate the active region of promoter by stepwise deletion. After that, I amplificate the promoter sequence, then construct the reporter gene plasmids. then transfect HepG2 cells by transient transfection. Next, I detect the activity of these reporter gene plasmids. Then the active region of promoter of HCBP6 is determined. At last,We predict that which transcriptional factor can bind with this active region,then check the prediction by electrophoretic mobility shift assay (EMSA).Results1.We have amplificated the promoter sequences successfully.2.We have constructed the reporter gene plasmids. 3.We find that there exist an binging site of Spl in the sequence of the promoter of HCBP6 by the bioinformatics.4.Transcriptional factor sp1 plays the key role in the regulation at transcriptional level by EMSA.Conclusions So I conclude that Sp1 is one of the key transcriptional factors of HCBP6.