近江牡蛎等7种养殖鱼虾贝类参照基因β-肌动蛋白cDNA序列的克隆与比较分析

为进一步研究水产养殖动物功能基因的表达调控,采用RT-PCR及RACE法,分离、克隆了近江牡蛎(Crassostrea ariakensis)肝脏β-肌动蛋白基因cDNA全序列.结果表明,近江牡蛎β-肌动蛋白基因cDNA全长1343bp,其中5′、3′非翻译区(UTR)分别长68bp、144bp,开放阅读框(ORF)为1131bp,编码376个氨基酸.实验同时成功获得了中华圆田螺(Cipangopaludina cahayensis)、日本沼虾(Macrobrachium nipponensis)、斑鳢(Channa maculata)、胡子鲶(Clarias fuscus)、鳙鱼(Aristichthys nobilis)、鲮鱼(Cirrhinus molitorella)6种重要淡水养殖动物肝脏β-肌动蛋白基因cDNA的核心序列及氨基酸序列.所获基因与其它动物类群的β-肌动蛋白基因氨基酸同源性高达96%以上,表明软体类、节肢类、鱼类、两栖类、鸟类和哺乳类等不同类群动物β-肌动蛋白基因在生物进化过程中高度保守.系统发育分析显示软体类、节肢类、鱼类、两栖类、鸟类和哺乳类脊椎动物β-肌动蛋白分类与传统的分类基本一致.

Cloning and Analysis of the Beta-Actin Gene in Jinjiang Oyster (Crassostrea ariakensis) and Other Six Cultivated Aquatic Animals

In order to study the expression and regulation of functional genes in aquatic animals, full-length cDNA of beta-actin gene in the liver of Jinjiang oyster (Crassostrea ariakensis) was obtained by RT-PCR and RACE methods. Results showed that the complete beta-actin cDNA in Jinjiang oyster was 1343bp in length, containing an open reading frame of 1131bp (encoding 376 amino acids), flanked by 68bp 5'UTR and 144bp 3'UTR. Partial cDNA sequences of mudsnail (Cipangopaludina cahayensis), shrimp (Macrobrachium nipponensis), Taiwan snakehead (Channa maculata), catfish (Clarias fuscus), bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella) beta-actin gene were also obtained. Sequence analysis revealed that the identities of beta-actin amino acid sequence among Jinjiang oyster and other species were more than 96%. This suggested that beta-actin was highly conserved. The phylogenetic tree constructed on the basis of beta-actin nucleotide sequence described accurately the relationship among mollusca, arthropod, fish, amphibian, bird and mammal.