烟粉虱内共生菌传毒相关groEL基因的克隆及其原核表达  

Cloning and prokaryotic expression of a viral binding protein gene (groEL) from endosymbionts in Bemisia tabaci

作　　者：谭周进[1] 谢丙炎[1] 肖启明[2] 杨宇红[1] 冯兰香[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]湖南农业大学植物保护学院,长沙410128

出　　处：《植物病理学报》2004年第4期314-318,共5页Acta Phytopathologica Sinica

基　　金：国家重点基础研究发展规划项目 ( 2 0 0 2CB1114 0 0 ) ;农业部农作物病虫草害生物防治资源研究与利用重点开放实验室开放基金 (LOBCR 2 0 0 3 0 1)

摘　　要：烟粉虱内共生菌 groEL基因编码一种 6 3kD的GroEL蛋白 ,利用一对特异性引物 ,通过PCR对长期培养在黄瓜上的烟粉虱体内 groEL基因进行了扩增 ,序列测定表明其长度为 16 6 8bp ,编码 5 5 5个氨基酸 ,与GenBank中烟粉虱内共生菌 groEL(AF130 4 2 1)的核苷酸同源性为 99.5 8%,仅 7个核苷酸发生了变异 ,二者氨基酸序列同源性为 99.2 8%,仅 4个氨基酸有别。构建了一个原核表达载体 ,表达得到了GroEL, a 63 kD viral binding protein encoded by groEL gene from endosymbionts in Bemisia tabaci, plays an important role in transmission of plant viruses. A groEL gene from endosymbionts in B. tabaci on cucumber was amplified by PCR method and sequenced. The nucleotide sequence indicated that groEL had 1 668 bp nucleotides and encoded 555 amino acids. This gene had 99.58% similarity on nucleotides level and 99.28% similarity on amino acid level to AF130421, a groEL gene from endosymbionts in B. tabaci in GenBank. There were only 7 variation sites in nucleotides and 4 variation sites in amino acids. SDS-PAGE showed a clear 76 kD fusion protein when the groEL gene was inserted into Pinpoint^(TM) Xa-1 T vector and expressed in E.coli JM109.

关 键 词：烟粉虱 内共生细菌 传毒相关基因 groEL基因 基因克隆 原核表达 蚜虫 

分 类 号：S433.3[农业科学—农业昆虫与害虫防治]