" Study on the cleavage of potato lea/roll virus cost protein gene in
vitro by ribozyme

GUO Xu-Dong; HA Shi-A-Gu-La; ZHANG He-Ling

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April 1997

GUO Xu-Dong, HA Shi-A-Gu-La and ZHANG He-Ling. ' Study on the cleavage of potato lea/roll virus cost protein gene invitro by ribozyme[J]. Virologica Sinica, 1997, 12(2).

Citation: GUO Xu-Dong, HA Shi-A-Gu-La, ZHANG He-Ling. " Study on the cleavage of potato lea/roll virus cost protein gene in vitro by ribozyme .VIROLOGICA SINICA, 1997, 12(2) : 149.

应用核酶体外切割马铃薯卷叶病毒外壳蛋白基因的研究

1.
内蒙古大学生物系

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。
- 核酶马铃薯卷叶病毒外壳蛋白基因体外转录体外切割

" Study on the cleavage of potato lea/roll virus cost protein gene in vitro by ribozyme

1.
Dep. of Biology, Inner Monglia

Abstract

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.
- RiboZyme Potato leafroll virus Coat protein gene Transcription in vitro Cleavage

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

" Study on the cleavage of potato lea/roll virus cost protein gene in vitro by ribozyme

1. Dep. of Biology, Inner Monglia

Abstract: 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.