党参肌动蛋白基因片段的克隆及序列分析  

Cloning and sequence analysis of actin gene fragment from roots of Codonopsis pilosula

作　　者：吴永娜[1] 李剑[1] 许瑞[2] 王引权[3] 张延红[3] 王惠珍[3] 张金林[1] 

机构地区：[1]兰州大学草地农业科技学院农业部草地农业生态系统重点开放实验室,甘肃兰州730020 [2]甘肃中医学院护理系,甘肃兰州730000 [3]甘肃中医学院药学系,甘肃兰州730000

出　　处：《中草药》2011年第12期2518-2522,共5页Chinese Traditional and Herbal Drugs

基　　金：甘肃省中医药科研立项课题(GZK-2010-41);兰州市科技发展计划项目(2010-1-39);教育部新世纪优秀人才支持计划;中央高校基本科研业务费专项资金项目(LZUJBKY-2010-2)

摘　　要：目的对党参肌动蛋白(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,以党参根部总RNA为模板,采用RT-PCR的方法扩增actin基因片段并连接到pMD19-T Simple载体上,阳性克隆经PCR检测后进行测序。结果得到一段603 bp的序列,序列分析表明,该片段编码200个氨基酸,与高等植物actin基因核苷酸序列同源性在78%以上,与其他肌动蛋白氨基酸序列同源性达90%以上。结论首次从党参中克隆出了actin基因,为有效利用该基因奠定了基础。Objective To clone and sequence the cDNA encoding actin gene from the roots of Codonopsis pilosula.Methods Degenerate primers were designed based on the conserved sequences of the actin gene from other plants.Total RNA was extracted from the roots of C.pilosula.Actin gene fragment was obtained by reverse transcription polymerase chain reaction（RT-PCR） and PCR products are sub-cloned into pMD19-T Simple vector.The positive clone identified by PCR was sequenced.Results The sequencing result revealed that the actin gene fragment from the roots of C.pilosula contained about 603 bp,encoding 200 amino acids.Sequence analysis suggested that the nucleotide sequence and the translated amino acid sequence shared over 78% and 90% of homology respectively with actin gene sequences from other higher plants.Conclusion It is the first report that a novel actin gene is cloned from roots of C.pilosula.This work lays a foundation for application to actin gene.

关 键 词：党参 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]