WANG Su-Yan, CHENG Gang, LIU Wei-Ping, XIE Yin and TIAN Bo. Construction of rice expression vector including antisense ribozyme and partial sequence of repli- case in rice dwarf virus[J]. Virologica Sinica, 1996, 11(2).
Citation: WANG Su-Yan, CHENG Gang, LIU Wei-Ping, XIE Yin, TIAN Bo. Construction of rice expression vector including antisense ribozyme and partial sequence of repli- case in rice dwarf virus .VIROLOGICA SINICA, 1996, 11(2) : 164.

抗水稻矮缩病毒的反义核酶及复制酶部分序列的合成、克隆及其水稻表达载体的构建

  • 采用反转录-聚合酶链式反应方法(RTP-CR),在人工合成的引物引导下,扩增出水稻矮缩病毒基因组第一片段(S1)全长序列及第五片段的部分序列(SSⅢ).将扩增的片段分别克隆到克隆载体pGEM7Zf(+)及pUC19的smal位点上,并进行了序列测定。在此基础上,利用pCR引入的方法将核酶序列引入到S5Ⅲ片段反义链上以构成反义核酶基因S5ⅢR,将S1片段5'端部分序列(S1-1)及S5ⅢR基因克隆到植物表达载体pROKⅡ上,构建成水稻转化载体pROK-S1-1'及pROK-S5ⅢR。

Construction of rice expression vector including antisense ribozyme and partial sequence of repli- case in rice dwarf virus

  • he whole sequence of RDV S1 fragment and partial sequence(S5Ⅲ)of RDV S5 fragmentwere synthesized and amplified by the method of reversed transcription-polymerase chain reaction(RT-PCR).Each amplified product was then cloned into pGEM7Zf(+)or pUC19 at the Smalsite,and sequenced.Based on these results,a hammer-head ribozyme sequence was introduced intoantisense chain of S5Ⅲfragment to construct antisense ribozyme gene S5ⅢR,Rice transformationvector pROK-S1-1' and pROK-S5ⅢRwere constructed by inserting S5ⅢRgene and 5'-end se-quence of S1 fragment(S1-1')into plant expression vector pROK Ⅱ.

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    Construction of rice expression vector including antisense ribozyme and partial sequence of repli- case in rice dwarf virus

    • 1. Institute of Microbiology ,Chinese Academy of Sciences

    Abstract: he whole sequence of RDV S1 fragment and partial sequence(S5Ⅲ)of RDV S5 fragmentwere synthesized and amplified by the method of reversed transcription-polymerase chain reaction(RT-PCR).Each amplified product was then cloned into pGEM7Zf(+)or pUC19 at the Smalsite,and sequenced.Based on these results,a hammer-head ribozyme sequence was introduced intoantisense chain of S5Ⅲfragment to construct antisense ribozyme gene S5ⅢR,Rice transformationvector pROK-S1-1' and pROK-S5ⅢRwere constructed by inserting S5ⅢRgene and 5'-end se-quence of S1 fragment(S1-1')into plant expression vector pROK Ⅱ.

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