Cloning and Expression in E ．coli of Nucleocapsid Protein Gene of Rice Grassy Stunt Virus

ZHANG Chun-Mei; TUN Jie-Jian; LIN Ai-Yang; XIE Lian-Hui

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April 2000

ZHANG Chun-Mei, TUN Jie-Jian, LIN Ai-Yang and XIE Lian-Hui. Cloning and Expression in E ．coli of Nucleocapsid Protein Gene of Rice Grassy Stunt Virus[J]. Virologica Sinica, 2000, 15(2): 200-203.

Citation: ZHANG Chun-Mei, TUN Jie-Jian, LIN Ai-Yang, XIE Lian-Hui. Cloning and Expression in E ．coli of Nucleocapsid Protein Gene of Rice Grassy Stunt Virus .VIROLOGICA SINICA, 2000, 15(2) : 200-203.

水稻草矮病毒核衣壳蛋白基因克隆及在大肠杆菌中的表达

1.
福建农业大学植物病毒研究所,福州350902

摘要

水稻草状矮化病于70年代曾在南亚、东南亚大面积发生,给当地的水稻生产造成严重损失,在我国也有分布[1]。其病原是水稻草矮病毒(Ｒｉｃｅｇｒａｓｓｙｓｔｕｎｔｖｉｒｕｓ,ＲＧＳＶ),为纤细病毒属(Ｔｅｎｕｉｖｉｒｕｓ)的一个成员,病毒粒体丝状,由核衣壳蛋白和基因组Ｒ?..
- 水稻草矮病毒
- , 核衣壳蛋白基因
- , 表达
- , 融合蛋白

Cloning and Expression in E ．coli of Nucleocapsid Protein Gene of Rice Grassy Stunt Virus

1.
Institute of Plant viiro[ogy，Fujian Agricutturat Universityz,Fuzhou 350002，China

Abstract

Based on the known RNA sequence of rice grassy stunt virus , IRRI isolate (RGSV IR), the cDNA of nucleocapsid protein (NCP) gene was obtained by RT PCR, with genomic RNAs of Shaxian isolate (RGSV SX) as template. The cDNA was then cloned into pGEM T vector and sequenced. The results showed that the nucleotide sequence between the two isolates was 99.4% identical. A bacterial expression plasmid pGTNCP which produced a fusion protein with molecular weight of about 62?kD was constructed using the cDNA clone and vector pGEX 2T. Western blot analysis showed that the fusion protein reacted strongly with antibodies raised against RGSV particles.
- Rice grassy stunt virus
- , Nucleocapsid protein gene
- , Expression
- , Fusion protein

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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

Cloning and Expression in E ．coli of Nucleocapsid Protein Gene of Rice Grassy Stunt Virus

1. Institute of Plant viiro[ogy，Fujian Agricutturat Universityz,Fuzhou 350002，China

Abstract: Based on the known RNA sequence of rice grassy stunt virus , IRRI isolate (RGSV IR), the cDNA of nucleocapsid protein (NCP) gene was obtained by RT PCR, with genomic RNAs of Shaxian isolate (RGSV SX) as template. The cDNA was then cloned into pGEM T vector and sequenced. The results showed that the nucleotide sequence between the two isolates was 99.4% identical. A bacterial expression plasmid pGTNCP which produced a fusion protein with molecular weight of about 62?kD was constructed using the cDNA clone and vector pGEX 2T. Western blot analysis showed that the fusion protein reacted strongly with antibodies raised against RGSV particles.