异色瓢虫HSP67B2基因的克隆及诱导表达分析

Molecular cloning and expression in different induction of Heat Shock Protein 67B2 gene in Harmonia axyridis (Pallas)

小分子热激蛋白是一种重要的分子伴侣,可以在环境胁迫下帮助蛋白质折叠和转运。本研究在转录组获得异色瓢虫Harmonia axyridis HSP67B2基因部分序列的基础上,通过RACE技术克隆得到了HSP67B2基因的cDNA全长序列(基因登录号:KJ155728),全长626 bp,其中编码140个氨基酸,3'非翻译区为81 bp,5'非翻译区为122 bp,软件分析预测显示该基因编码蛋白的分子量为15.97 kD,理论等电点为6.61。采用荧光定量PCR技术对异色瓢虫HSP67B2基因在不同发育阶段、低温诱导条件和饥饿处理下的表达进行了研究,结果表明:HSP67B2基因在幼虫期的表达量最高;在低温诱导和饥饿处理下,HSP67B2基因出现上调表达并且表达效率显著提高,表明HSP67B2基因可以通过调控小分子热激蛋白的合成使异色瓢虫适应环境变化。

Small heat shock protein, an important molecular chaperone, can assist protein folding and transporting under environmental stress. Based on Harmonia axyridis transcriptome, the experiment got the full cDNA of HSP67B2 in H. axyridis which consists of 626 nucleotides （ GenBank accession number： KJ155728） including an nucleotides that encodes 140 amino acid residues and a 122 or 81 nucleotide for 5 ＇- or 3＇- non - coding region respectively. Software analysis predicts HSP67B2 has a molecular weight of 15.97 kD and electronic point of 6.61. Meantime real - time PCR was adopted to detect the expression of H. axyridis HSP67B2 at different developmental stages and under the treatment of cold induction and starvation. The results show that the expression level of HSP67B2 is highest in the larval stage and HSP67 B2 gene is significantly up - regulated under the treatment of cold induction and starvation. The results imply that HSP67B2 gene can make 1-1. axyridis adapt to enviorment changes by precisely the regulation of the small heat shock protein synthesis.