HCV特异性M1GS核酶的构建及体外切割活性研究  

Study on the in Vitro Cleavage Activity of an Artificial HCV-specific M1GS Ribozyme

作　　者：张文军[1] 宁容[1] 张欣[2] 李红枝[1] 

机构地区：[1]广东药学院基础学院,广州510006 [2]暨南大学生命科学技术学院,广州510632

出　　处：《中国生物工程杂志》2008年第9期99-103,共5页China Biotechnology

基　　金：广州市科技局科技计划项目(2005J1-C0291);广东药学院博士科研启动基金(43543106)资助项目

摘　　要：针对HCV基因组中较为保守的区域—5′UTR,设计一段GS引导序列,并与大肠杆菌RNase P的催化亚基—M1RNA的3′末端共价结合,构建序列特异性M1GS核酶—M1GS-HCV/C20。体外实验证实,所构建的人工核酶对HCV5′UTR具有明显的靶向切割活性,且这种切割发生于靶序列的特定位点。研究将为进一步阐明该核酶在胞内的活性、乃至动物模型内评价其抗病毒效果提供实验材料,从而为新型抗HCV药物及反义基因治疗的研究奠定基础。A sequence-specific M1GS ribozyme （M1GS-HCV/C20） has been successfully constructed by covalently linking an oligonucleotide （guide sequence, GS） to the 3＇ terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The engineered ribozyme is targeted to the most conservative sequence （5＇ UTR） of HCV genome, and can effectively cleave the substrate RNA segment in vitro. Undoubtly, the M1GS- HCV/C20 would be a useful experimental material to futher study its cleavage activity in vivo, and can be even used for evaluating its anti-viral effect in the animal model. It was believed that the study would markedly facilitate the research of a general gene targeting agent for anti-HCV applications, and layed the foundation for developing a new nucleic acid drug and a novel strategy of anti-HCV therapy.

关 键 词：核酶P 引导序列 丙型肝炎病毒 5’非编码区 

分 类 号：Q789[生物学—分子生物学]