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当前位置: 首页 > 文章 > 棉铃虫保幼激素环氧水解酶基因的克隆及其原核表达 环境昆虫学报 2009,31 (2) 114-123
Position: Home > Articles > Cloning of cDNA-encoding juvenile hormone epoxide hydrolase in Helicoverpa armigera(Hbner) and its expression in E. coli Journal of Environmental Entomology 2009,31 (2) 114-123

棉铃虫保幼激素环氧水解酶基因的克隆及其原核表达

作  者：
宁长明;吴孔明;高希武;程红梅
单  位：
中国农业大学昆虫系;中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室;中国农业科学院生物技术研究所
关键词：
基因克隆;表达;Western blot;保幼激素;保幼激素环氧水解酶;棉铃虫
摘  要：
通过兼并引物PCR结合RACE技术,克隆了棉铃虫Helicoverpa armigera(Hbner)保幼激素环氧水解酶(juven-ile hormone epoxide hydrolase,JHEH)的基因,该基因的开放阅读框为1389bp,编码463个氨基酸。预测蛋白分子量为52kD,等电点为6.39。N末端存在由20个氨基酸组成的疏水性信号肽序列,存在组成JHEH催化三联体的氨基酸Asp(227)、Glu(403)和His(430)以及组成阴氧离子洞的氨基酸Tyr(298)、Tyr(373)和HGWP花样结构。其氨基酸序列与其它鳞翅目昆虫有很高的同源性。在大肠杆菌中表达JHEH基因的编码区,Western blot结果表明,棉铃虫JHEH已被成功表达。利用该基因序列可以在分子水平上研究棉铃虫JHEH基因的时空表达情况,进一步研究保幼激素(juvenile hormone,JH)的代谢途径及其功能。
译  名：
Cloning of cDNA-encoding juvenile hormone epoxide hydrolase in Helicoverpa armigera(Hbner) and its expression in E. coli
作  者：
NING Chang-Ming1,2,WU Kong-Ming2,GAO Xi-Wu1,CHENG Hong-Mei3 (1. Department of Entomology,China Agricultural University,Beijing 100193,China; 2. State Key Laboratory for Biology of Plant Disease and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China; 3. Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China)
关键词：
gene cloning; expression; Western blot; juvenile hormone; juvenile hormone epoxide hydrolase; Helicoverpa armigera
摘  要：
The cDNA-encoding juvenile hormone epoxide hydrolase (JHEH) was cloned by degenerative PCR combined with RACE techniques in Helicoverpa armigera (Hbner). The open reading frame of JHEH was 1389 bp ,encoding 463 amino acid residues. Its predicted molecular weight was 52kD and isoelectric point was 6.39. A hydrophobic signal sequence with 20 amino acids in N-terminal region,the residues Asp(227),Glu(403),His(430) that form the potential catalytic triad,and the residues Tyr(298),Tyr(373),and the HGWP motif that consist of the potential oxyanion hole were found in the deduced amino sequences. The H.armigera JHEH amino acid sequences have high homology to JHEH from other lepidoptern species. The coding region of JHEH was expressed in E.coli,and Western blot analysis showed that the JHEH fusion protein was successfully expressed. This will not only facilitate further study on its spatio-temporal expression pattern at molecular level,but also be useful to the understanding of the metabolic pathway and the functions of juvenile hormone (JH).

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