当归肌动蛋白基因片段的克隆及序列分析  

Cloning and sequence analysis on Actin gene fragment from Angelica sinensis

作　　者：吴永娜[1] 胡静[1] 王引权[2] 李剑[1] 张金林[1] 

机构地区：[1]兰州大学草地农业科技学院,甘肃兰州730020 [2]甘肃中医学院药学系,甘肃兰州730000

出　　处：《中草药》2012年第12期2485-2489,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(81060327);甘肃省中医药科研立项课题(GZK-2010-41);教育部新世纪优秀人才支持计划(NCET-11-0217)

摘　　要：目的对当归肌动蛋白(Actin)基因进行克隆及序列分析。方法根据已经克隆的植物Actin基因的保守序列设计一对简并性引物,以当归根部总RNA为模板,采用RT-PCR方法扩增Actin基因片段并连接到pMD19-T载体上,阳性克隆经PCR检测后进行测序。结果得到一段598 bp的序列,序列分析表明,该片段编码198个氨基酸,与高等植物Actin基因核苷酸序列同源性在83%以上,与其他肌动蛋白氨基酸序列同源性达94%以上。结论首次从当归中克隆出了Actin基因,为有效利用该基因奠定了基础。Objective Cloning and sequence analysis on the cDNA fragment encoding Actin gene in the roots of Angelica sinensis.Methods A pair of degenerate primers were designed based on the conservative sequences of the cloned Actin gene from other plant species.Taking total RNA from the roots of A.sinensis as template,Actin gene fragment was obtained by reverse transcription polymerase chain reaction（RT-PCR） and PCR products were sub-cloned to pMD19-T vector.The identified positive clone was sequenced.Results The result revealed that the Actin gene fragment from the roots of A.sinensis contained 598 bp,encoding 198 amino acids.Sequence analysis suggested that the nucleotide sequence shared over 83% of homology with Actin gene from other higher plants,and the amino acid homology with other Actin reached over 94%.Conclusion It is the first report that a novel Actin gene is cloned from the roots of A.sinensis.This work lays a base for the effective application of Actin gene.

关 键 词：当归 肌动蛋白 基因克隆 序列分析 RT-PCR 

分 类 号：R282.2[医药卫生—中药学]