广东药学院学报2017，Vol.33Issue(1)：103-107,5.DOI:10.16809/j.cnki.2096-3653.2016111601
AIV PB2基因靶向性M1GS核酶的构建及其体外切割活性测定
Construction and cleavage activity in vitro of a M1GS ribozyme that targeting to the PB2 gene of avian influenza virus
摘要
Abstract
Objective To construct a MIGS ribozyme that targeting to replicase PB2 gene of avian influenza virus ( AIV) , and provide a reference for development of the novel anti-AIV strategy. Methods Based on the ribonuclease P ( RNase P) derived from Escherichia coli, a guide sequence ( GS) complementary with PB2 gene sequence was designed, and covalently linked to the 3′end of catalytic subunit of RNase P ( M1 RNA) by PCR. Then the activity of this targeting M1GS ribozyme was measured by the in vitro cleavage assay. Results Two kinds of M1GS ribozymes, namely M1GS-C341 and M1GS-T436 , were constructed. The ribozyme M1GS-T436 specifically cleaved the target RNA, however the ribozyme M1GS-C341 showed the nonspecific cleavage activity. Conclusion A potential M1GS ribozyme ( M1GS-T436 ) , that possess the specific cleavage activity, has been successfully constructed, which may lay the foundation for further study of its intracellular and in vivo anti-viral activity.关键词
M1GS核酶/禽流感病毒/PB2基因/抗病毒Key words
M1GS ribozyme/AIV/PB2 gene/antiviral分类
生物科学引用本文复制引用
李小英,伍婧茹,刘琳,张文军..AIV PB2基因靶向性M1GS核酶的构建及其体外切割活性测定[J].广东药学院学报,2017,33(1):103-107,5.基金项目
广东省自然科学基金项目(S2012010009471) （S2012010009471）
病毒学国家重点实验室开放基金项目(2012008) （2012008）
