Effect of specific nucleotide G11 on cleavage activity of hairpin ribozymes in vitro

被引:0
|
作者
Song, YH
Lin, JS
Kong, XJ
Li, B
Liu, NZ
Jin, YX [1 ]
机构
[1] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China
[2] Huazhong Univ Sci & Technol, Tongji Med Coll, Tongji Hosp, Inst Liver Dis, Wuhan 430030, Peoples R China
关键词
hairpin ribozyme; kinetic of enzyme; point mutant;
D O I
暂无
中图分类号
T [工业技术];
学科分类号
08 ;
摘要
To study the effect of specific G(11) nucleotide on cleavage activity of hairpin ribozyme in vitro, the P-32-labeled pKC transcript (267 nt) containing hepatitis B virus core region is used as target RNA. Synthetic hairpin ribozyme genes with G(11) mutational site are cloned into ribozyme vector p1.5 to create pHpRz for preparation of ribozyme. Non-labeled HpRz transcripts are incubated with P-32- labeled target-RNAs under different conditions and autoradiographed after gel electrophoresis. The results show that the cleavage activity of hairpin ribozyme is closely related to the matching of the bases between hairpin ribozyme and the substrate. The G(11) site is not essential to cleavage activity of hairpin ribozyme in vitro, so the selection of target sequence matching G(11) site of hairpin ribozyme is not limited to this site.
引用
收藏
页码:345 / 349
页数:5
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