黄粉甲储存蛋白hexamerin基因的克隆及表达分析

Cloning and Expression Analysis of Storage Protein hexamerin genes in Tenebrio molitor

基于RACE技术从黄粉甲蛹中克隆得到储存蛋白hexamerin基因(TmolHex1和TmolHex2),应用生物信息软件和荧光定量PCR技术分析了基因的序列和表达特征。结果表明:克隆获得的TmolHex1和TmolHex2基因的开放阅读框分别为2133bp和2103bp,分别编码710和700个氨基酸,其编码蛋白预测理论分子量分别为85.53kDa和84.35kDa,等电点分别为6.05和6.72。TmolHex1和TmolHex2氨基酸序列中存在信号肽序列,富含芳香氨基酸,且具有芳香储存蛋白家族特征序列。TmolHex1和TmolHex2基因在黄粉甲的老熟幼虫和蛹脂肪体中表达量最高,表明其可能在生长发育、变态和能量代谢方面发挥重要作用。

The storage protein hexamerin genes(TmolHex1 and TmolHex2) were cloned from pupae of Tenebrio molitor by RACE technology. Their sequences were analyzed by bioinformatic softwares and their expression patterns were analyzed using the real-time fluorescence quantitative PCR. The cloned TmolHex1 and TmolHex2 genes respectively contain open reading frames of 2 133 and 2 103 bp, which encode 710 and 700 amino acid residues. Their predicted molecular weights of amino acid sequences are 85.53 kDa and 84.35 kDa,and isoelectric points are 6.05 and 6.72, respectively. Sequence analysis revealed that there are signal peptide sequences in the amino acid sequences of TmolHex1 and TmolHex2. They are rich in aromatic amino acids, and have the characteristic sequences of aromatic storage protein family. TmolHex1 and TmolHex2 genes show the highest expression levels at the mature instar larvae and in pupal fat body of T. molitor, indicating that they may play important roles in development, metamorphosis, and energy metabolism.