GS介导RNase P对人巨细胞病毒ul54基因mRNA的切割作用  

Targeted Cleavage of the Human Cytomegalovirus ul54 Gene mRNA by M1GS

作　　者：李弘剑[1] 黎景光[1] 苏运贞[1] 陈浩军[1] 李月琴[1] 叶飞舟[1] 张欣[1] 周天鸿[1] 

机构地区：[1]暨南大学生命科学技术学院,广东广州510632

出　　处：《病毒学报》2006年第4期275-281,共7页Chinese Journal of Virology

基　　金：国家自然科学基金(编号:30370776);广东省自然科学基金重大项目(编号:36703);广东省自然科学基金(编号:021162;000718)

摘　　要：引导序列(Guide Sequences,GSs)是与mRNA靶序列互补并引导RNase P切割的小RNA片段。设计与人巨细胞病毒HCMV(Human Cytomegalovirus,HCMV)ul54基因D片段mRNA序列互补的GS,将其共价结合到大肠杆菌来源RNase P催化核心M1 RNA,构建成T7-M1GS核酶。通过对ul54基因D片段转录产物体外切割实验和将T7-M1GS构建在含有U6启动子的逆转录病毒载体,与构建在真核载体pEGFP-N1的ul54基因D片段共转染人宫颈癌细胞系HeLa的体内切割实验,证实该核酶具备对ul54基因D片段mRNA的特异切割能力,为利用核酶治疗HCMV感染提供实验基础。Guide sequences （GSs） are small RNA molecules that consist of a sequence complementary to a targeted mRNA and render the targeted RNA susceptible to degradation by the catalytic RNA subunit of RNase P from Escherichia coli （M1 RNA）. A sequence-specific ribozyme （T7-MIGS） derived from GSs and M1 RNA was used to target a region of μ/54 gene D segment mRNA of human cytomegalovirus （HCMV）.Through the experiments about targeted cleavage in vitro and in vivo of the HCMV μ/54 gene D segment mRNA by T7- MIGS, the results showed that T7-MIGS can effectively cleave μ/54 mRNA. This provides an experimental basis for treatment of HCMV by ribozymes.

关 键 词：人巨细胞病毒HCMV μ/54基因 RNASE P 引导序列(GSs) 荧光定量PCR 

分 类 号：R373.9[医药卫生—病原生物学]