棉铃虫HaKettin1基因的全长cDNA克隆、序列分析和表达谱

【目的】为了克隆棉铃虫Helicoverpa armigera编码肌肉蛋白Kettin基因的全长cDNA序列以及鉴定该基因在棉铃虫发育周期内的表达模式。【方法】利用兼并引物，通过分段RT-PCR和5′-和3′-RACE的方法克隆全长cDNA序列。利用半定量RT-PCR进行表达谱分析。【结果】编码棉铃虫Kettin蛋白的基因HaKettin1全长cDNA序列为13 805 bp，包含一个13 365 bp的开放阅读框，编码4 454个氨基酸，蛋白分子量约为504.3 kD。组织表达结果显示HaKettin1基因在棉铃虫的整个生育周期都有表达，幼虫期的表达尤为显著。【结论】HaKettin1与家蚕的Kettin蛋白具有90％的同源性，表明鳞翅目昆虫的Kettin蛋白之间具有很高的保守性。表达谱结果显示HaKettin1基因在棉铃虫的整个发育过程中都发挥重要作用。

cDNA cloning, sequence analysis and expression profiling of HaKettin1 gene from Helicoverpa armigera

Objective] To clone the full length cDNA sequence of Kettin gene from Helicoverpa armigera and analyze its expression profiles in the life cycle of H.armigera development.Methods] Using multiple RT-PCR and 5'-,3'-RACE to clone the full length cDNA sequence of HaKettin1,and semi-quantitative RT-PCR to analyze the expression profiles.Results] The full length of HaKettin1 cDNA sequence is 13 805 bp,containing an open reading frame of 13 365 bp.The deduced amino acid sequence consists of 4 454 amino acids,with molecular mass about 504.3 kD.The results of expression profiles showed HaKettin1 was expressed in all the cycle of insect life,and the expression of HaKettin1 at larval stage was higher than at other developmental stages.Conclusions] Percent identity of HaKettin1 protein and Bombyx mori Kettin protein is 90%,suggesting that Kettin proteins are much conserved among Lepidoptera insects.The results of expression profiles suggest that HaKettin1 plays an important role in the development of H.armigera.