农业科学与技术：英文版
農業科學與技術：英文版
농업과학여기술：영문판
Agricultural Science ＆ Technology
2011年
7期
971-974
,共4页
尚常花%朱顺妮%袁振宏%王忠铭
尚常花%硃順妮%袁振宏%王忠銘
상상화%주순니%원진굉%왕충명
巴夫杜氏藻%β-肌动蛋白基因%克隆
巴伕杜氏藻%β-肌動蛋白基因%剋隆
파부두씨조%β-기동단백기인%극륭
Dunaliella parva%β-actin gene%Cloning
[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。
[目的]穫得巴伕杜氏藻β-肌動蛋白基因cDNA全長序列。[方法]以巴伕杜氏藻cDNA為模闆,採用簡併引物進行PCR擴增,穫得533 bp特異cDNA片段。在此基礎上,設計特異引物,採用5′-GenomeWalking和3′-RACE的方法,穫得基因的5′-耑DNA序列和3′-耑cDNA序列,進而穫得β-肌動蛋白基因cDNA全長序列。[結果]穫得瞭巴伕杜氏藻β-肌動蛋白基因的特異cDNA片段、5′-耑DNA和3′-耑cDNA片段。經拼接後,擴增齣全長cDNA。β-肌動蛋白基因cDNA全長1 754 bp,包括1 137 bp的開放讀碼框和617 bp的3′-非翻譯區序列。氨基痠序列相似性分析髮現,巴伕杜氏藻β-肌動蛋白氨基痠序列與杜氏鹽藻、萊茵衣藻等的同源性較高。繫統髮育分析錶明,巴伕杜氏藻β-肌動蛋白與杜氏鹽藻的相似性最高。[結論]首次穫得瞭巴伕杜氏藻β-肌動蛋白基因cDNA全長序列併髮現巴伕杜氏藻β-肌動蛋白基因非常保守。
[목적]획득파부두씨조β-기동단백기인cDNA전장서렬。[방법]이파부두씨조cDNA위모판,채용간병인물진행PCR확증,획득533 bp특이cDNA편단。재차기출상,설계특이인물,채용5′-GenomeWalking화3′-RACE적방법,획득기인적5′-단DNA서렬화3′-단cDNA서렬,진이획득β-기동단백기인cDNA전장서렬。[결과]획득료파부두씨조β-기동단백기인적특이cDNA편단、5′-단DNA화3′-단cDNA편단。경병접후,확증출전장cDNA。β-기동단백기인cDNA전장1 754 bp,포괄1 137 bp적개방독마광화617 bp적3′-비번역구서렬。안기산서렬상사성분석발현,파부두씨조β-기동단백안기산서렬여두씨염조、래인의조등적동원성교고。계통발육분석표명,파부두씨조β-기동단백여두씨염조적상사성최고。[결론]수차획득료파부두씨조β-기동단백기인cDNA전장서렬병발현파부두씨조β-기동단백기인비상보수。
[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.
