灰飞虱共生菌groEL基因的克隆、生物信息学分析及其植物双元表达载体构建  

Clonging and Bioinformatic Analysis of groEL from Wolbachia Endosymbiont in Small Brown Planthopper and Establishment of Plant Binary Expression Construct

作　　者：曹卿[1,2] 杨杰[1] 王军[1] 范方军[1] 仲维功[1] 张玉琼[2] 张云华[2] 

机构地区：[1]江苏省农业科学院粮食作物研究所 江苏省优质水稻工程技术研究中心 国家水稻改良中心南京分中心,南京210014 [2]安徽农业大学生命科学院,合肥230036

出　　处：《分子植物育种》2010年第1期35-40,共6页Molecular Plant Breeding

基　　金：国家科技支撑计划重大项目(2006BADO2A03);国家转基因生物新品种培育科技重大专项(2009ZX08001-005B,019);江苏省自然科学基金项目(BK2009321);江苏省政府留学奖学金;江苏省农业科技自主创新基金项目(CX[07]603);江苏省农业科学院基金项目(6110703;6510806)共同资助

摘　　要：本研究克隆了江苏南京地区灰飞虱(Laodelphax striatellus Fallén)体内共生菌Wolbachia的groEL基因全长序列。序列分析表明,其开放阅读框为1659bp,编码552个氨基酸残基,分子量为58.884kD,等电点为5.01。序列比对发现克隆的groEL基因与已登录的groEL基因(登录号:EF468716)起始密码子下游424bp处存在一个核苷酸的差异,与另一groEL基因(登录号:DQ356890)完全一致。利用PROTEIN DATA BANK在线软件对灰飞虱的groEL蛋白立体结构进行预测,结果表明灰飞虱的groEL蛋白为同型寡聚复合物,具有分子伴侣功能。本研究进一步构建了含该groEL基因和潮霉素基因的双元表达载体,为下一步转化水稻奠定了基础。In this paper, we cloned a Wolbachia gene groEL from small brown planthopper （Laodelphax striatellus Fallen） collected in Nanjing, Jiangsu Province. Sequence analysis showed that the cloned fragment contained an open reading frame of 1 659 bp, encoding 552 amino acids resides with a calculated molecular weight of 58.884 kD and isoelectric point of 5.01. Sequence alignment demonstrated that the obtained groEL sequence was completely matched one groEL （Accession No. DQ356890）, and one SNP was found 424 bp downstream of initiation codon of another reported groEL （accession No. EF 468716）. Moreover, we predicted the three dimensional structure of groEL protein by online software PROTEIN DATA BANK, and found that it was a homo-oligomeric complex with the function of molecular chaperone. Futrhermore, we established a binary transgenic vector haboring the cloned groEL gene and hygromyein resintant gene （HPT）, which would facilitate further rice transformation.

关 键 词：灰飞虱 共生菌 WOLBACHIA GROEL 生物信息学分析 分子伴侣 植物双元表达载体 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]