The Preliminary Study Of Interaction Between HCV Core Protein And Homo Sapiens Pancreatic Colipase Protein

background:Hepatitis C virus (HCV) chronic infection can lead to chronic liver inflammation, necrosis and fibrosis, some patients can develop liver cirrhosis and even hepatocellular carcinoma(HCC).Substantial statistical data show that HCV infection and liver steatosis, diabetes and other metabolic diseases have a correlation.The liver steatosis is one of the histology features of the chronic hepatitis C (CHC), which suggested that CHC is also a kind of metabolic diseases.HCV infection can lead to metabolic syndrome(MS) ,including Liver steatosis,insulin resistance and so on. Pancreatic cell secretory protein is closely related to glucose and lipid metabolism. Studies have shown that the interaction between HCV core protein and the host cell plays an important role in liver steatosis.Objective:To further study the interaction between HCV core protein and the host pancreatic cell.Methods:1.We transformed yeast cell expression vector pGBKT7-core into yeast strain AH109 and performed yeast two hybrid by mating AH109 containing HCV core gene with Y187 containing plasmids of human pancreas cDNA library and screened of proteins binding to HCV core protein from human pancreas cDNA library.2.Homo sapiens colipase gene was amplified with reverse transcription polymerase chain reaction(RT-PCR).3. We constructed eukaryotic expression vector pcDNA3.1-myc-his-clps and pcDNA3.1(-)core and confirmed the interaction between HCV core protein and Homo sapiens colipase protein using co-immunoprecipitation and Western blotting. 4.We constructed eukaryotic expression vector pACT-clps and pBIND-core and confirmed the interaction between HCV core protein and Homo sapiens colipase protein using mammalian two-hybrid experiment.Results:1.We found eleven proteins which interact with HCV core protein.2. Homo sapiens pancreatic colipase gene was obtained .3.We constructed eukaryotic expression vector pcDNA3.1-myc-his-clps and confirmed the intra-cellular interaction between HCV core protein and homo sapiens colipase protein.4.We constructed eukaryotic expression vector pACT-clps and confirmed the intra-cellular inter action between HCV core protein and homo sapiens colipase protein.Conclusion:We found eleven proteins which could bind to HCV core protein and confirmed the interaction between HCV core protein and homo sapiens colipase protein in intra-ce llular , which laid the foundation for further studying the mechanism of HCV core protein.