用cDNA末端扩增法克隆柞蚕攻击素(attacin)基因及序列分析

利用cDNA末端扩增(RLM-RACE)方法,克隆了柞蚕攻击素(attac in)基因cDNA序列(GenBank登陆号:AY960680)。柞蚕攻击素基因cDNA序列全长912 bp,其中699 bp的蛋白质编码区可编码233个氨基酸。预测蛋白质分子量为25 kD,等电点(pI)为7.54。柞蚕攻击素基因中包含2个内含子。S ignal P 3.0 Server程序分析结果显示,柞蚕攻击素第1-17位氨基酸为信号肽序列。蛋白质功能区分析预测,柞蚕攻击素在第47-112氨基酸之间为攻击素-N(attac in-N)功能区,第113-233位氨基酸之间为攻击素-C(attac in-C)功能区。以邻接法(ne igh-bor-join ing,NJ)建立了与其它昆虫攻击素的系统关系图。

Cloning and Analysis of cDNA of the Attacin Gene From the Chinese Oak Silkworm, Antheraea pernyi, Using RLM-RACE Method

The cDNA sequence of a attacin gene(GenBank accesion No.AY960680)from Chinese Oak Silkworm,Antheraea pernyi,was determined using RLM-RACE method.The full-length cDNA is(912 bp)with a(699 bp) open reading frame(ORF),which encodes a protein of 233 amino acids.The protein is about 25 kD and its pI value is 7.54.From genomic DNA,there are two introns and three exons within the open reading frame.Amino acid sequence analysing by Signal(P 3.0) Server showed that there is a signal peptide of 17 amino acids at the N-terminal.Prediction of protein founctional domain indicated that the N-terminal domain of Ap-attacin is from 47 to 112 amino acid and C-terminal domain is from 113 to 233 amino acid by Pfam.In addition,an amino acid phylogenetic tree of attacins among different insects was constructed by using the neighbor-joining method.