The Verification Of Interaction Between HCBP6 And HCVcore

Objective:To testify the interaction between HCBP6 and HCVcore by CO-IP. And to understand the subcellular localization of HCBP6 and HCVcore by LSCM(laser scan confocal microscopy). To identify the function of HCBP6 is useful to establish search foundation.Methods:1.Real-time PCR was adopted to compare different quantitative expression of HCBP6 from 7 cell lines.The cell line with higher expression was selected for subsequent experiment to evalute interaction between proteins on eukaryotic cell level.2.HepG2 cells was transfected with pcDNA3.1-hcvcore,pcDNA 3.1/myc-his(-)-hcbp6,pcDNA3.1and pcDNA3.1/myc-his(-), sepa-rately. The interaction between HCBP6 and HCVcore was evaluated by CO-IP.3.The GFP expressing vector of HCBP6 and pDsRed expressing vector of HCVcore were constructed after confirming by DNA sequence analysis and restriction enzyme digestion. The plasmids were transfected into HepG2 cell lines by lipofectamine, respectively. Subcellular localization was detected by the LSCM (laser scan confocal microscopy).Results:1.The expression of HCBP6 in different cell lines from highest to low-est is Capan2,HepG2,Hela,Huh7,7721,7701 and 7402.2.Both co-transfection and independent transfection of pcDNA 3.1-hcvcore and pcDNA3.1/myc-his(-)-hcbp6 to HepG2 are proved to be successful.3.The GFP expressing vector of HCBP6 and pDsRed expressing vector of HCVcore were successful expressing in the HepG2 cell lines. They are both detected in cytoplasm.Conclusion:1.HCVcore is the hepatitis C virus core protein.To study the role with HCBP6 should choose one between the hepatocellelar carcinoma cell lines.The expression of HCBP6 in HepG2 cell line is high and HepG2 is one kind of hepatoma carcinoma cell lines. HepG2 cell line was chosen for the following experiment.2.Interaction between HCBP6 and HCVcore are proved.3.Both HCBP6 and HCVcore are detected in cytoplasm. Subcellular localization establish the premise for interaction between HCBP6 and HCVcore.HCBP6 educe biologic activity in cytoplasm.