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虾夷扇贝β-actin基因的克隆和序列分析
引用本文:傅立元,高祥刚,李云峰,王健,赫崇波.虾夷扇贝β-actin基因的克隆和序列分析[J].生物技术通报,2010(4).
作者姓名:傅立元  高祥刚  李云峰  王健  赫崇波
作者单位:1. 辽宁师范大学生命科学学院,大连,116029
2. 辽宁省海洋水产科学研究院,辽宁省海洋水产分子生物学重点实验室,大连,116023
基金项目:辽宁省科学技术计划重大重点项目,辽宁省海洋与渔业科研计划项目,国家海洋公益性行业科技专项 
摘    要:为进一步研究虾夷扇贝功能基因的表达调控。利用SMART cDNA文库构建试剂盒成功构建了健康虾夷扇贝外套膜和肾脏两种组织的cDNA文库。对随机选取的4009个克隆进行5′端测序,比对,筛选出1条β肌动蛋白同源序列,对此EST序列两端进行扩增、测序,得到肌动蛋白基因cDNA全长序列。肌动蛋白基因cDNA全长1536bp(不包括polyA),5′端非编码区84bp,3′端非翻译区321bp,阅读框1131bp,编码377个氨基酸。在基因组DNA中,该基因被一个内含子分为两段,内含子位于第41和第42个氨基酸之间,长度为1498bp。系统发育分析显示该肌动蛋白属于β类型。本研究得到的虾夷扇贝β-肌动蛋白基因可以被用于作为定量某种虾夷扇贝mRNA的标准,这为继续研究虾夷扇贝其它功能基因,及其分子生物的进一步研究、促进其他相关分子发育和系统进化研究奠定了基础。

关 键 词:虾夷扇贝  Cdna文库  β-肌动蛋白  基因克隆  序列分析  β-actin  gene

Cloning and Characterization of β-actin Gene from Mizuhopecten yessoensis
Fu Liyuan,Gao Xianggang,Li Yunfeng,Wang Jian,He Chongbo.Cloning and Characterization of β-actin Gene from Mizuhopecten yessoensis[J].Biotechnology Bulletin,2010(4).
Authors:Fu Liyuan  Gao Xianggang  Li Yunfeng  Wang Jian  He Chongbo
Institution:Fu Liyuan1,2Gao Xianggang2Li Yunfeng2Wang Jian2He Chongbo2(1College of Life Sciences,Liaoning Normal University,Dalian 116029,2Liaoning Ocean , FisheryScience Institute,Liaoning Key Laboratoty of Marine Fishery Molecular Biology,Dalian 116023)
Abstract:The cDNA library from mantle and kidney of Mizuhopecten yessoensis was constructed using Creator SMART cDNA construction kit and large numbers of colonies were randomly picked and sequenced from the 5'. One EST with high homology with β-actin gene of other species was found and then the complete express sequence of β-actin from mizuhopecten yessoensis was obtained by PCR. The cDNA of this gene was 1 536 bp with 1 131 bp of ORF, which encodes 177 amino acids. The intron of this gene was 1 498 bp which cited between the 41st amino and the 42nd amino. The gene in this study is used for other genes' quantitative expression of Mizuhopecten yessoensis as an intemal control.
Keywords:Mizuhopecten yessoensis  cDNA library  Gene clone  Sequence analysis
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