Cloning and Characterization of β-actin Gene in Dunaliella parva  

巴夫杜氏藻β-肌动蛋白基因的克隆和分析(英文)

作　　者：尚常花[1] 朱顺妮[1] 袁振宏[1] 王忠铭[1] 

机构地区：[1]中国科学院广州能源研究所中国科学院可再生能源与天然气水合物重点实验室,广东广州510640

出　　处：《Agricultural Science & Technology》2011年第7期971-974,共4页农业科学与技术（英文版）

基　　金：Supported by Comprehensive Strategic Cooperation Project betweenGuangdong Province and Chinese Academy of Sciences (2010A0-90100010);National Key Technology R&D Program of the 12thFive-year Plan of China (2011BAD14B03);Foundation of KeyLaboratory of Renewable Energy and Natural Gas Hydrate,ChineseAcademy of Sciences (y107j6)~~

摘　　要：[Objective] The aim was to obtain the full-length cDNA sequence of Dunaliella parva β-actin gene.[Method] Based on the highly conserved amino acid regions of known β-actin,a pair of degenerate primers was synthesized to amplify 533 bp cDNA sequences in Dunaliella parva.Then,the 5＇ genomic DNA and 3＇ cDNA sequences were obtained by Genome walking and 3＇-RACE technology based on the obtained sequence.According to the sequences of the 5＇-termini and 3＇-termini,specific primers were synthesized to obtain the full-length cDNA.[Result] The full-length β-actin cDNA included 1 137 bp open reading frame（ORF）,617 bp of 3＇ noncoding region.Similarity analysis indicated that the highest similarity was found between Dunaliella parva and Dunaliella salina.The Dunaliella parva β-actin also showed wide similarity with other algae.[Conclusion] The full-length cDNA sequence of D.parva was firstly obtained,which was highly conserved.[目的]获得巴夫杜氏藻β-肌动蛋白基因cDNA全长序列。[方法]以巴夫杜氏藻cDNA为模板,采用简并引物进行PCR扩增,获得533 bp特异cDNA片段。在此基础上,设计特异引物,采用5′-GenomeWalking和3′-RACE的方法,获得基因的5′-端DNA序列和3′-端cDNA序列,进而获得β-肌动蛋白基因cDNA全长序列。[结果]获得了巴夫杜氏藻β-肌动蛋白基因的特异cDNA片段、5′-端DNA和3′-端cDNA片段。经拼接后,扩增出全长cDNA。β-肌动蛋白基因cDNA全长1 754 bp,包括1 137 bp的开放读码框和617 bp的3′-非翻译区序列。氨基酸序列相似性分析发现,巴夫杜氏藻β-肌动蛋白氨基酸序列与杜氏盐藻、莱茵衣藻等的同源性较高。系统发育分析表明,巴夫杜氏藻β-肌动蛋白与杜氏盐藻的相似性最高。[结论]首次获得了巴夫杜氏藻β-肌动蛋白基因cDNA全长序列并发现巴夫杜氏藻β-肌动蛋白基因非常保守。

关 键 词：Dunaliella parva β-actin gene CLONING 

分 类 号：Q943.2[生物学—植物学]