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

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ＲＮＡ组成2]。基因组含六个ｓｓＲＮＡ片段,均为双义编码3,4],其中ＲＮＡ5的毒义互补链编码核衣壳蛋白(ｎｕｃｌｅｏｃａｐｓｉｄｐｒｏｔｅｉｎ,ＮＣＰ)3]。本文报道应用ＲＴＰＣＲ技术获得ＲＧＳＶ沙县分离株(ＲＧＳＶＳＸ)ＮＣＰ基因的ｃＤＮＡ克隆,并得到其在大肠杆…

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

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.