摘要
针对马铃薯卷叶病毒外壳蛋白基因第356～358位点“GUC”．设计、合成了一种“锤头状”核酶。将核酶基因克隆在体外转录载体PSPT19的SP6启动子下游；同时将PLRVCPcDNA亚克隆在体外转录载体pSPT18的SP6启动子下游。利用SP6RNA聚合酶分别体外转录，获得核酶分子和靶RNA序列。在41℃保温进行核酶切割反应，检测到预期大小且被切开的两个RNA短片段。
A hammerhead structure ribozyme was designed and synthesized to potato leafroll virus Chinese isolate (PLRV - Ch) genome within 356 to 358 nt 'GUC' in coat protein (CP) gene. DNA sequence encoding the ribozyme was inserted into the position downstream from SP6 promoter of in vitro transcription vector pSPT19. The target PLRV CP gene cDNA was subcloned in vector pSPT18. The RNA molecules of riboZyme and target Sequence were generated by transcription with SP6 RNA polymerase in vitro. After cleavage reaction at 41℃, two.expected short RNA fragments were observed.
出处
《中国病毒学》
CAS
CSCD
1997年第2期149-154,共6页
Virologica Sinica
基金
国家自然科学基金
关键词
马铃薯卷叶病毒
核酶
外壳蛋白基因
体外切割
RiboZyme,Potato leafroll virus,Coat protein gene,Transcription in vitro,Cleavage
