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| 象耳豆根结线虫Hsp70基因的克隆与原核表达 | |
| 引用本文： | 练冬梅,王会芳,赵志祥,陈绵才.象耳豆根结线虫Hsp70基因的克隆与原核表达[J].植物病理学报,2015,45(1):7-13. |
| 作者姓名： | 练冬梅 王会芳 赵志祥 陈绵才 |
| 作者单位： | 1.海南省农科院农业环境与植物保护研究所/海南省植物病虫害防控重点实验室,海口 571100; 2.海南大学环境与植物保护学院,海口 571101 |
| 基金项目： | 国家自然科学基金项目(31260424);公益性行业(农业)科研专项(201103018);“十二五”国家科技支撑计划项目(2012BAD19B06-07) |
| 摘 要： | 采用RT-PCR、c DNA末端快速扩增法(RACE),克隆了象耳豆根结线虫Hsp70基因的全长c DNA(Me Hsp70)序列。Me Hsp70 c DNA全长2 203 bp,含有1 959 bp的开放阅读框,编码653个氨基酸,相对分子量为71.09 k Da,具有3段Hsp70家族的签名序列,Gen Bank登录号KF739434。同源性分析表明,氨基酸序列与其他真核生物的Hsp70序列具有很高的相似性。该Hsp70与其他物种中的Hsp70进行系统进化分析,结果显示,Hsp70的系统发育树不能体现物种间的亲缘关系,推测其反映的是不同物种间Hsp70生物学功能的相似性程度。构建了一个原核表达载体p EASY-E1-Me Hsp70,当IPTG终浓度为0.4~1.0 mmol/L时,能诱导表达融合蛋白。Me Hsp70基因的克隆和表达,将为象耳豆根结线虫的生态适应性机理研究提供依据。 |
| 关 键 词： | 象耳豆根结线虫 Hsp70 克隆 表达 |
Cloning and prokaryotic expression of Hsp70 from Meloidogyne enterolobii |
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| LIAN Dong-mei,WANG Hui-fang,ZHAO Zhi-xiang,CHEN Mian-cai.Cloning and prokaryotic expression of Hsp70 from Meloidogyne enterolobii[J].Acta Phytopathologica Sinica,2015,45(1):7-13. | |
| Authors: | LIAN Dong-mei WANG Hui-fang ZHAO Zhi-xiang CHEN Mian-cai |
| Institution: | 1 Institute of Environment & Plant Protection, Hainan Academy of Agricultural Sciences, Hainan Key Laboratory for Control of Plant Diseases and Insect Pests , Haikou 571100, China; 2 Environment and Plant Protection College, Hainan University, Haikou 571101, China |
| Abstract: | The full length cDNA (MeHsp70) of Hsp70 from Meloidogyne enterolobii was cloned by using RT-PCR, rapid amplification of cDNA ends ( RACE) . The full-length cDNA of MeHsp70 was 2 203 bp containing an open reading frame (ORF) of 1 959 bp encoding a polypeptide of 653 amino acids with a predicted molecular mass of 71.09 kDa, which carried three important and intact Hsp70 signature sequences, which was registered in GenBank with accession No. KF739434. The result of sequence similarity-analysis revealed that the translated molecules showed high homology to other eukarya. The Hsp70s phylogenetic analysis showed that the phylogenetic tree could not reflect the genetic relationship of the organisms, maybe similarity of biological functions was revealed. The complete ORF encoding MeHsp70 was inserted into pEASY-E1 and an expression vector pEASY-E1-MeHsp70 was constructed. The MeHsp70 protein was induced to express by 0. 4-1.0 mmol/L IPTG. This study provided theoretical basis supporting research on mechanisms of the ecological adaptability of M. enterolobii. |
| Keywords: | Meloidogyne enterolobii Hsp |
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