池蝶蚌细胞色素P450 cDNA及其蛋白的序列分析

Analysis of the cDNA cequence and protein structure of cytochrome P450 from Hyriopsis schlegelii

在实验室已获得的池蝶蚌(Hyriopsis schlegelii)部分细胞色素P450基因序列的基础上,运用RACE PCR的方法获得池蝶蚌细胞色素CYP基因(定义为Hs-CYP450)的全长cDNA。运用相关生物软件和在线软件对HsCYP450基因序列以及其蛋白的一、二、三级结构进行了初步分析。结果显示,Hs-CYP450基因全长1 809bp,ORF框1 431bp,共编码476个氨基酸;氨基酸组成成分中亮氨酸(Leu)含量最多,约9.5%,而最少的则是色氨酸,仅0.8%;通过疏水性分析显示,该蛋白属亲水性可溶性蛋白;二级结构预测显示该蛋白α螺旋(H)结构含量较多,约占46.22%,并均匀分布在蛋白中;β折叠(E)相对较少,约9.87%左右,多分布在两端。建立的系统进化树结果表明,Hs-CYP450基因序列与其他动物的细胞色素CYP基因有较高的同源性,在众多物种中和太平洋牡蛎(Crassostrea gigas)同源性相对较高。

In this study,Hs-CYP450 was cloned from by the method of RACE PCR.Then,relevant biologi-cal softwares and online softwares were used to make a preliminary analysis of Hs-CYP450 gene sequence, as well as the primary,secondary and tertiary structures of this protein.Our results indicated that the Hs-CYP450 cDNA present a full length of 1809bp that consisted of an open reading frame （ORF）of 1 431 bp encoding a protein of 476 amino acids,The content of leucine （Leu）accounted for 9.5% in the protein,but tryptophan content was only 0.8%;The hydrophobicity analysis showed that this protein was a hydrophilic soluble protein;The prediction of secondary structure suggested thatαhelix （H）content was 46.22% and distributed evenly in the protein;βfold （E）was about 9.87% and was located at both ends.The results of phylogenetic tree showed that Hs-CYP450 gene sequence had a high homology with other animals CYP gene,and the Crassostrea gigas shared the highest among these species.