中山大学学报自然科学版 ›› 2010, Vol. 49 ›› Issue (2): 72-78.

• 研究论文 • 上一篇    下一篇

异色瓢虫的HSP 90基因的克隆与特性分析

唐 斌1,2,王世贵1,王峰巍2,3,庞 虹3,张 帆2   

  1. (1杭州师范大学杭州市动物适应与进化重点实验室,浙江 杭州 310036;2北京市农林科学院植保环保所,北京 100089;3中山大学生命科学学院,广东 广州 510275)
  • 收稿日期:2009-02-28 修回日期:1900-01-01 出版日期:2010-03-25 发布日期:2010-03-25

Cloning and Characterization Analysis of Heat Shock Protein 90 Gene from Harmonia axyridis( Pallas)

TANG Bin1,2, WANG Shigui1, WANG Fengwei2,3, PANG Hong3, ZHANG Fan2   

  1. (1. Hangzhou Key Laboratory of Animal and Evolution,Hangzhou Normal University, Hangzhou 310036, China;2. Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100089, China;)3. School of Life Sciences, Sun Yatsen University, Guangzhou 510275, China
  • Received:2009-02-28 Revised:1900-01-01 Online:2010-03-25 Published:2010-03-25

摘要: 采用同源克隆和锚定PCR技术, 从异色瓢虫 Harmonia axyridis (Pallas)中克隆到热休克蛋白HSP 90 基因的cDNA全序列(Genbank number: FJ501962)。cDNA全长2 480 bp,包含3′非编码区域(UTR)为200 bp和5′UTR为126 bp,开放阅读框(ORF) 长2.154 bp,编码717个氨基酸。预测的相对分子质量为82.230, 理论等电点为4.96,无糖基化位点、跨膜结构和信号肽。在N端具有 HSP 90 基因保守的ATPase结构,含有HSP90家族的C末端的保守序列EEVD。与赤拟谷盗 Tribolium castaneum 相比较,同源性高达90%,系统发育分析也表明两者的亲缘关系最近。 HaaHSP 90 基因的克隆和比较分析为进一步深入研究异色瓢虫的抗逆机理及其进化具有重要意义。

关键词: 异色瓢虫, HSP 90 , 克隆, 特性分析, 序列分析

Abstract: The heat shock protein 90 (HSP 90) gene from Harmonia axyridis (Pallas) was cloned using techniques of homological cloning and anchored PCR. The full length of cDNA sequence is 2 480 bp(Genbank number: FJ501962), containing a 3′UTR (untranslated region) of 200 bp, a 5′UTR of 126 bp, and an ORF of 2 154 bp which encodes a polypeptide of 717 amino acids with an estimated molecular weight of 82 230 and p I of 4.96. There are no glycosylation sites, putative transmembrane domain and signature peptide. It contains ATPase conservative domain of (HSP90) in Nsite and EEVD signature sequence on Csite. Compared with Tribolium castaneum, they are 90% identities and Phylogenetic analysis shows that these two species are most closely related. Cloning and comparison analysis of HSP 90 genes from Harmonia axyridis (Pallas) could be useful in the studies of antistress mechanism, tolerance improvement breeding and evolution for these animals.

Key words: Harmonia axyridis, HSP 90, cloning, characterization analysis, sequence analysis

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