HCV核心蛋白与HCBP1在哺乳双杂交系统中的相互作用.doc

HCV核心蛋白与HCBP1在哺乳双杂交系统中的相互作用
作者:陈云茹刘敏陈天艳张树林陈瑞琳张曦石磊
【摘要】目的:探讨HCVcore与HCBP1的相互作用. 方法:PCR分别扩增含HCV core及HCBP1的cDNA片段,克隆入pGEM T载体,经测序正确后再分别克隆入哺乳双杂交的表达质粒pM 和pVP16中,并转染入COS7细胞,48 h后检测细胞中报告基因CAT的表达水平. 结果:CATELISA检测显示pMcore与pVP16HCBP1实验孔A405 ,高于其它阴性对照,但低于阳性对照组A值. 结论:哺乳双杂交系统证实HCV核心蛋白与新基因HCBP1有一定的相互作用,可进一步研究HCBP1的细胞功能.
【关键词】 HCV核心蛋白;哺乳双杂交;报告基因
基金项目:国家自然科学基金(30471532);陕西省科学技术研究发展计划项目(2004K15G1)

【Abstract】 AIM: To analyze the interaction between HCV core protein and HCBP1 using mammalian twohybrid assay. METHODS: cDNA fragments encoding HCV core protein and HCBP1 were amplified by PCR and subsequently cloned into pGEM T vector, respectively. After verified by sequencing, they were respectively subcloned into two hybrid plasmids, pM and pVP16. Then, pM
core, pVP16HCBP1 and pG5CAT, a reporter vector, all were cotransfected into COS7 cells. Fortyeight hours later, the interaction between HCV core protein and HCBP1 was assayed by detecting CAT gene expression. RESULTS: CATELISA showed that the A value of the cotransfection experiment group was , significantly higher than those of the other negative control groups and lower than that of the positive control group. CONCLUSION: Mammalian twohybrid assay confirms HCBP1 can bind with HCV core protein to some extent. Based on this, we can study the function of HCBP1 further.
【Keywords】 hepatitis C virus