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当前位置: 首页 > 文章 > 蕙兰Actin基因的克隆及序列分析 河南农业科学 2013,42 (6) 107-111
Position: Home > Articles > Molecular Cloning and Sequence Analysis of Actin Gene from Orchid(Cymbidium faberi) Journal of Henan Agricultural Sciences 2013,42 (6) 107-111

蕙兰Actin基因的克隆及序列分析

作  者：
田云芳;刘艺平;李涵;苏金乐
单  位：
河南农业大学林学院;濮阳职业技术学院生物工程系
关键词：
蕙兰;肌动蛋白基因;克隆;序列分析
摘  要：
持家基因Actin常被用作定量、半定量PCR试验的内参。为研究其他基因的调控机制或外源基因在蕙兰中的相对表达提供内参,根据GenBank已经登录的肌动蛋白(Actin)基因的同源核苷酸保守序列,设计简并引物,利用RT-PCR的方法,以蕙兰花蕾期的花葶为材料,克隆了4个Actin基因片段。序列分析结果表明,4个Actin基因片段长度均为1 062bp,编码354个氨基酸,具有Actin基因的标记位点:肌动蛋白(YVGDEAQs.KRG和WISKaEYDE)和肌动蛋白类似物(LLTEApLNPkaNR)。各片段之间同源性高达95.97%,经BLAST分析,与文心兰同源性可达94%。其氨基酸序列与萼脊兰(AED94091.1)和蝴蝶兰(AAF71265.1)同源性达99%。得到的4个基因序列是Actin基因的同源片段,分别命名为CfACT1、CfACT2、CfACT3和CfACT5,并在GenBank注册,登录号分别为JN177718、JN177719、JN177720和JN177721。
译  名：
Molecular Cloning and Sequence Analysis of Actin Gene from Orchid(Cymbidium faberi)
作  者：
TIAN Yun-fang1,2,LIU Yi-ping1,LI Han3,SU Jin-le1(1.College of Forestry of Henan Agricultural University,Zhengzhou 450002,China; 2.Department of Life Science of Zhengzhou Normal University,Zhengzhou 450044,China; 3.Bioengineering Department of Puyang Vocational and Technical College,Puyang 457000,China)
关键词：
Cymbidium faberi;Actin;cloning;sequence analysis
摘  要：
Housekeeping gene Actin is often used as an internal control gene of RT-PCR and qRT-PCR.In order to study the molecular regulatory mechanisms of other genes and relative expression level of exogenous gene,a pair of primers was designed according to the conserved sequences of the Actin gene in GenBank.A PCR-based homologous cloning strategy was used to identify Actin genes from the pedicels of Cymbidium faberi.Four Actin genes were successfully cloned and characterized.The sequencing result revealed that each of four Actin gene fragments contained 1 062 bp,encoding a protein of 354 amino acids.Each Actin contained the Actin family signature sequence(YVGDEAQs.KRG and WISKaEYDE) and Actin-related proteins signature sequence(LLTEApLNPkaNR).The fragments shared 95.97% nucleotide sequence homology with each other and 94% nucleotide sequence homology with Oncidium hybrida ACT2(JN981137).The deduced amino acid sequence shared 99% homology with Actin of Sedirea japonica(AED94091.1) and Phalaenopsis(AAF71265.1).The cloned genes were Actin gene fragments,named as CfACT1,CfACT2,CfACT3 and CfACT5,respectively.They were registered into GenBank(accession number:JN177718,JN177719,JN177720 and JN177721).

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