曼氏无针乌贼(Sepiella maindroni)β-肌动蛋白基因的cDNA全长克隆与序列分析  

CLONING AND ANALYSIS OF THE FULL-LENGTH cDNA SEQUENCE OF SEPIELLA MAINDRONI β-ACTIN GENE

作　　者：李继姬[1] 郭宝英[1] 吴常文[1] 

机构地区：[1]浙江海洋学院海洋科学学院浙江省海洋养殖装备与工程技术重点实验室,舟山316004

出　　处：《海洋与湖沼》2011年第6期787-793,共7页Oceanologia Et Limnologia Sinica

基　　金：国家高技术研究发展计划(863计划)项目"名贵头足类苗种规模繁育关键技术";2010AA10A404号;国家海洋公益性行业科研专项;201005013号;国家自然科学基金项目;31101937号;2011年浙江省大学生科技创新(新苗人才计划)项目;2011.05-2012.03

摘　　要：采用RT-PCR和快速扩增cDNA末端(rapid amplification of cDNA ends,RACE)技术首次克隆了曼氏无针乌贼β-肌动蛋白基因的cDNA全序列,该序列全长为2000bp,由长197bp的5’非翻译区(untranslated region,UTR),669bp的3’非翻译区,和1134bp的开放阅读框(open reading frame,ORF)组成。阅读框共编码377个氨基酸,推算的分子量约为42.0kDa,理论等电点为5.16。曼氏无针乌贼β-actin氨基酸序列中Ile12、Ser172、Ser174、Gln223、His227、Ile231、Gly232、Ser320、Glu328、Thr360等10个氨基酸残基具有特异性,以及2个特殊的氨基酸残基位点和2个软体动物特有的氨基酸残基。曼氏无针乌贼β-actin氨基酸序列与软体动物、节肢动物、脊椎动物的相似性高达97%。NJ法系统进化分析显示曼氏无针乌贼首先与软体动物聚在一起,然后与节肢动物聚在一起,再与鱼类、两栖类、哺乳类聚在一起。In this paper, a 2000bp full-length cDNA sequence of β-actin gene from Sepiella maindroni was obtained with RT-PCR and rapid amplification of cDNA ends （RACE） technique. It consists of a 197bp 5＇ untranslated region （UTR）, an 1134bp open reading frame （ORF） and a 669bp 3＇UTR. The translated protein is composed of 377 amino acids, with 42.0kDa molecular weight, and its calculated isoelectric point was 5.16. The amino acid sequence of J3-actin in Sepiella maindroni has ten specific amino acid residues： Ile12, Ser172, Ser174, Gln223, His227, Ile231, Gly232, Set320, Glu328 and Thr360, respectively. In addition, two other particular base sites of amino acid residues were found in these sequences. Similarly, two characteristic amino acid residues of some mollusks were obtained. The amino acids sequence of β-actin in Sepiella rnaindroni shared the high similarity with Molluscs, Arthropod and Vertebrate animals （97%）. Neighbor-Joining （NJ） tree suggested that Sepiella maindroni clustered with Mollusca firstly, and then clustered with Arthropoda, finally clustered with Fish, Amphibians, and Mammals.

关 键 词：曼氏无针乌贼 CDNA 快速扩增cDNA末端(RACE) β-肌动蛋白基因 

分 类 号：Q24[生物学—细胞生物学]