两种核酶对烟草花叶病毒RNA片段的体外剪切  

TWO RIBOZYMES THAT CLEAVE TOBACCO MOSAIC VIRUS RNA FRAGMENTS IN VITRO

作　　者：于善谦[1,2] 吴政宏[1,2] 朱乃硕[1,2] 秦进[1,2] 赵立山[1,2] 章道道[1,2] 

机构地区：[1]复旦大学微生物系 [2]复旦大学化学系

出　　处：《病毒学报》1997年第1期79-82,共4页Chinese Journal of Virology

基　　金：国家科委攀登计划

摘　　要：两种核酶对烟草花叶病毒ＲＮＡ片段的体外剪切于善谦吴政宏朱乃硕秦进赵立山章道道（复旦大学微生物系，上海２００４３３）关键词核酶，烟草花叶病毒，体外剪切核酶（Ｒｉｂｏｚｙｍｅ）是具有特殊结构的ＲＮＡ分子，表现酶解活性，对核酸的切割有顺序专一性。...According to two targets at 5505 and 5954 within the regions known to encode the TMV vulgare strain 30kD protein and coat protein genes,two hammerhead ribozymes Rz-1,Rz-2 and their substrates Sb-1,Sb-2 were designed and synthesized.Their cDNA were inserted into pGEM-3Z plasmid under the control of T7 promoter.The Escherichia coli TG2 transformed by recombined plasmids pGRz-1,pGSb-1,pGRz-2,pGSb-2 and pGRzSb-2 that is a cis-splicing system between Rz-2 and Sb-2 were selected and identified by selective medium with Amp-Xgal-IPTG.The ribozyme and the substrate RNAs were synthesized by transcription by T7 polymerase in vitro.The catalytic reaction of the ribozyme to its substrate was done in the TE buffer solution which contained Mg 2+ and spermine at 25℃,The results of the reaction showed that the substrate Sb-1 RNA of 34nt was cleaved by Rz-1 into two RNA fragments of 20nt and 14nt when the ratio of the concentration of Rz-1 to Sb-1 reached to 20:1. Similar result was also observed in the reaction between Sb-2 and Rz-2 when the ratio of the concentration of Rz-2 to Sb-2 reached to 16:1.RzSb-2 cis-splicing system showed higher cis-acting self-cleaving activity.Self-cleaving reaction of the RzSb-2 showed much higher activity when the ribozyme Rz-2 was added into the cis-splicing system.The effect of some factors on the efficiency of ribozyme activity isdiscussed.

关 键 词：植物病毒 核酶 烟草花叶病毒 RNA 体外剪切 

分 类 号：S432.41[农业科学—植物病理学]