水稻矮缩病毒基因组分析与部分基因片段的表达及功能研究

水稻矮缩病毒基因组分析与部分基因片段的表达及功能研究

李毅, 徐洪, 程明非, 郑宏红, 冒志娟, 肖锦, 张福建, 明小天, 曲林, 刘一飞, 李玮, 赵晓岚, 潘乃Sui, 陈章良

北京大学生命科学学院，蛋白质工程及植物基因工程国家重点实验室，北京，100871

收稿日期:1997-12-22 出版日期:1998-04-20 发布日期:1998-04-20

Functional Genomic Analysis of Rice Dwarf Virus

LI Yi, XU Hong, CHENG Mingfi, ZHENG Honghong, MAO Zhijun, XIAO Jin, ZHANG Fujian, MING Xiaotian, QU Lin, LIU Yifei, LI Wei, ZHAO Xiaolan, PAN Naisui, CHEN Zhangliang

The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing, 100871

Received:1997-12-22 Online:1998-04-20 Published:1998-04-20

摘要/Abstract

摘要： 根据本实验室对水稻矮缩病毒(Rice Dwarf Virus, RDV)中国福建分离株基因组全序列的测定，分析了其编码蛋白的功能，并将RDVS6，S9，S10，S11编码区cDNA分别克隆到表达载体pTrcHisA, pBV221, pTrcHisB和pBV221中，构建成表达质粒pTH-S6, pBVP9, pTH-S10, pBVP11。分别用IPTG或温度诱导大肠杆菌DH5α，经SDS-PAGE及相应抗体的Western blotting检测证明S6, S9, S10, S11编码蛋白在大肠杆菌中获得成功表达。另外，还将RDV外层外壳蛋白基因S8、微核心蛋白基因S7及编码非结构蛋白基因S9通过基因枪导入到水稻植株中，抗病性正在研究之中。

关键词: RDV基因组分析, 原核表达, Westernblotting, 转基因水稻

Abstract: Rice dwarf virus full genome 12 gene segments were cloned and 25000 nucleotides were sequenced. The gene functions of each segment were analyzed according to amino acid sequences and experimental data. The coding regions of segments S6, S9, S10 and S11 encoding nonstractural proteins were cloned into expression vector pTricHisA, pBV221, pTric HisB, pBV221, respectively. The expressed products were analyzed by SDS-PAGE and further confirmed by Western-blotting. Using the E.coli expression products, the function of Pns11 was studied. The results showed that Pns11 is a ds-/ss-RNA and DNA binding protein and it may play very important roles in virus genome assortment or packaging. Gene segments S2, S7, S8 and S9 which encode virus most outer coat protein, minor core protein, outer coat protein and nonstructural protein were transferred into rice via biolistic bombardment and regenerated rice plants were obtained. The resistance to RDV infection of these transgenic rice plants are in testing.

Key words: ROV genome analysis, expression in E.coli, Western blotting, Transgenic plants

中图分类号:

Q939.46

李毅, 徐洪, 程明非, 郑宏红, 冒志娟, 肖锦, 张福建, 明小天, 曲林, 刘一飞, 李玮, 赵晓岚, 潘乃Sui, 陈章良. 水稻矮缩病毒基因组分析与部分基因片段的表达及功能研究[J]. 北京大学学报（自然科学版）.

LI Yi,XU Hong,CHENG Mingfi,ZHENG Honghong,MAO Zhijun,XIAO Jin,ZHANG Fujian,MING Xiaotian,QU Lin,LIU Yifei,LI Wei,ZHAO Xiaolan,PAN Naisui,CHEN Zhangliang. Functional Genomic Analysis of Rice Dwarf Virus[J]. Acta Scientiarum Naturalium Universitatis Pekinensis.

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https://xbna.pku.edu.cn/CN/Y1998/V34/I2/332