【摘要】 应用RT-PCR技术克隆了水稻瘤矮病毒(Rice gall dwarf virus,RGDV)广东分离物基因组的第10片段,并测定了全序列。结果表明,RGDV广东分离物S10(登录号EF532325)全长1 198 bp,含有一个ORF,编码一条由320氨基酸组成、推测分子量约36 kDa的多肽。与泰国分离物相应组分相比,基因结构基本一致,核苷酸和氨基酸序列同源性分别为96.2%和98.8%;S10编码多肽与水稻矮缩病毒(Rice dwarf virus,RDV)S9编码蛋白及伤瘤病毒(Wound tumor virus,WTV)S11编码蛋白也分别具有29%和33%的相似性。本研究还将S10 cDNA克隆至原核表达载体pET28b(+)上,通过IPTG诱导在大肠杆菌BL21(DE3)中得到了高效表达,并利用His. Bind树脂纯化得到电泳纯级制品。本工作为进一步研究S10编码蛋白的结构与功能奠定了一定的基础。更多还原

【Abstract】 The full length cDNA of S10 sequence of Rice gall dwarf virus(RGDV) Guangdong isolate was cloned into pMD18-T vector and the complete nucleotide sequence was determined.The results showed that S10 had a length of 1 198 nt encoding a polypeptide of 320 amino acids with a Mr of 36 kDa.Sequence comparison showed that the S10 of RGDV Guangdong isolate had the same genome organization with the Thailand isolate,and shared 96.2% nucleotide and 98.8% amino acid sequence identities.The predicted polypeptide of RGDV S10 also shared 29% and 33% identities respectively with the proteins encoded by Rice dwarf virus(RDV)S9 and Wound tumor virus(WTV) S11.The RGDV S10 cDNA was cloned to pET28b(+) and highly expressed in Escherichia coli BL21(DE3),and the expressed recombinant protein was purified by His.Bind column.These works will promote the structural and functional research of RGDV S10 protein.更多还原