大螟热激蛋白83基因克隆及序列分析
Molecular cloning and sequence analysis of the HSP83 gene in Sesamia inferens (Walker) (Lepidoptera: Noctuidae)
孙 猛** 陆明星 汤小天 杜予州
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DOI：10.7679/j.issn.2095-1353.2014.148
作者单位：扬州大学园艺与植物保护学院，扬州 225009
中文关键词：大螟，HSP83，基因克隆，序列分析
英文关键词：Sesamia inferens, HSP83, gene cloning, sequence analysis
中文摘要：
【目的】 近年来，大螟Sesamia inferens (Walker) 对水稻的为害逐渐加重并成为水稻的重要害虫之一。HSP83家族作为分子伴侣参与生物生长发育并对外界刺激产生响应，对生物功能蛋白质的正确折叠及胁迫信号传递有着重要意义。本研究旨在克隆大螟HSP90家族HSP83基因的cDNA及基因组全长序列，分析其基本特性。【方法】 应用RT-PCR及RACE技术从大螟中克隆HSP83基因的cDNA序列；获取基因组序列，进行基因组验证，并与cDNA序列比较分析其有无内含子；进行系统发育分析。【结果】 大螟HSP83基因被命名为Sihsp83（GenBank登录号：KM077137），长度为2 496 bp，开放阅读框长2 154 bp，编码717个氨基酸，推测分子量为82.6 ku。该基因编码的氨基酸序列中含有5个HSP90家族保守序列，在C-末端存在细胞质定位信号。大螟HSP83基因组序列长度为2 218 bp（GenBank登录号：KM077138），不含内含子。在系统发育树中，大螟HSP83与其他夜蛾科昆虫的HSP90家族聚为一支。【结论】 获得的大螟HSP83基因是HSP90基因家族成员；大螟的HSP83是细胞质热激蛋白，具有很高的保守性。
英文摘要：
[Objectives] The pink borer, Sesamia inferens (Walker) has in recent years become a major pest of rice crops that is responsible for steadily increasing crop losses. The heat shock protein 90 (HSP90) family is of great significance for the correct folding and transmission of proteins in organisms. HSP90 affects the growth and development of organisms and their responses to external stimuli. Our study’s aim was to clone the full length cDNA and genome sequences of the HSP83 gene from S. inferens, and analyze its basic characteristics. [Methods] RT-PCR and RACE were used to clone the full-length cDNA of HSC70 gene from S. inferens. The genomic sequence was determined by genomic validation, introns were analyzed by sequence analysis and a phylogeny constructed. [Results] The complete cDNA sequence was obtained and named as Sihsp83 (GenBank accession no.: KM077137). This was 2 496 bp in length and contained an opening reading frame (ORF) of 2 154 bp encoding 717 amino acid residues with a predicted molecular weight of 82.6 ku. Five conserved sequences of the HSP90 family exist in this amino acid sequence. The cytoplasmic localization signal was present in the C-terminal. Analysis of the genomic DNA sequence (GenBank accession no.: KM077138) demonstrated that the HSP83 gene does not contain introns. [Conclusion] Full-length cDNA and genome sequences of the HSP83 gene from S. inferens were obtained, and do not contain introns. Sequence analysis confirms that HSP90 is very conservative.