双齿围沙蚕CYP4基因的克隆及序列分析  

Cloning and sequence analysis of cytochrome CYP4 family in polychaete Perinereis aibuhitensis

作　　者：周一兵[1] 陈雪[1] 杨大佐[1] 万良[1] 王斌[1] 王丽丽[2] 孙静波 

机构地区：[1]大连海洋大学辽宁省海洋生物资源恢复与生境修复重点实验室,辽宁大连116023 [2]国家海洋环境监测中心,辽宁大连116023 [3]宝生物工程(大连)有限公司,辽宁大连116600

出　　处：《大连海洋大学学报》2011年第6期507-513,共7页Journal of Dalian Ocean University

基　　金：国家"863"计划项目(2006AA10Z410);国家海洋公益性行业科研专项(200805069);国家自然科学基金资助项目(30901107;41106115)

摘　　要：根据已知的绿沙蚕Nereis virens细胞色素氧化酶CYP342A1保守区设计引物,采用RACE技术首次从双齿围沙蚕Perinereis aibuhitensis体壁肌肉中克隆出细胞色素氧化酶CYP4基因全长序列。分析结果表明,该序列全长为1 857 bp,5'端非翻译区为186 bp,3'端非翻译区为225 bp,阅读框为1 446 bp,编码为481个氨基酸。第422～431氨基酸序列符合P450所具有的结构保守共有序列FxxGxxxCxG,第319～322氨基酸序列为K螺旋特征基序ExxR区,第282～294氨基酸序列含有CYP4家族特征序列EVDTFMFEGHDTT。经Blast与GenBank中已知物种的氨基酸序列进行同源性比对,CYP4与多毛类绿沙蚕Nereis virens CYP4BB1、CYP342A1(AY453408)氨基酸同源性为73%、36%,与多毛类小头虫Capitella capitata CYP4AT1(AY574044)同源性为39%。据此推断,双齿围沙蚕CYP4基因属于CYP4B亚家族。In the present study, the partial cDNA of CYP4 was amplified from the total RNA of epidermis muscle of the polychaete,Perinereis aibuhitensis, by reverse transcription with the primers based upon the CYP342A1 from Nereis virens. Then other primers were designed according to the partial gene of Perinereis aibuhitensis CYP4, and two fragments about 371 bp and 1044 bp were obtained by using 3＇ RACE and 5＇ RACE. Sequence analysis revealed a 1857 bp cDNA containing the 186 bp 5＇-untranslated region,225 bp 3＇ -untranslated regions and 1 446 bp open reading frame encoding 481 amino acid. The identity rate of deduced amino acid sequence of CYP4 was subjected to Blast searching in NCBI. The results showed that the CYP4 shared 73% identity with CYP4BB1,36% identity with CYP342A1 of Nereis virens and 39% identity with CYP4AT1 of Capitella capitate. The deduced amino acid sequence of CYP4 contained the conserved motifs of P450 family（FxxGxxxCxG） and had K-helix motifs of P450 family （ExxR）. In addition, it has the characteristic sequences of CYP4 family （EVDTFMFEGHDTT）. So it is in- ferred that the eDNA sequence might belong to the subfamily of CYP4B.

关 键 词：双齿围沙蚕 CYP4 RACE 生物信息学分析 

分 类 号：Q78[生物学—分子生物学]