CAJ | 학술논문

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4 009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1 536 bp(不包括poly A) , 5'端非编码区84 bp, 3'端非翻译区321 bp,阅读框1 131 bp,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1 498 bp.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.
위진일보연구하이선패공능기인적표체조공.이용SMART cDNA문고구건시제합성공구건료건강하이선패외투막화신장량충조직적cDNA문고.대수궤선취적4 009개극륭진행5'단측서,비대,사선출1조β기동단백동원서렬,대차EST서렬량단진행확증、측서,득도기동단백기인cDNA전장서렬.기동단백기인cDNA전장1 536 bp(불포괄poly A) , 5'단비편마구84 bp, 3'단비번역구321 bp,열독광1 131 bp,편마377개안기산.재기인조DNA중,해기인피일개내함자분위량단,내함자위우제41화제42개안기산지간,장도위1 498 bp.계통발육분석현시해기동단백속우β류형.본연구득도적하이선패β-기동단백기인가이피용우작위정량모충하이선패mRNA적표준,저위계속연구하이선패기타공능기인,급기분자생물적진일보연구、촉진기타상관분자발육화계통진화연구전정료기출.
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.