箭筈豌豆两个肌动蛋白基因片段的克隆及其序列分析.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 9 筈豌豆两个肌动蛋白基因片段的克隆及序列分析 张磊 刘志鹏 张吉宇 张妙青 王彦荣 (, 730000) : 通过同源克隆获得了箭筈豌豆两个不同的 Actin基因片段, 为研究该箭筈豌豆其它基因的表 状况提供了内 标参照根据近缘物种 Actin 因序列设计 一对引物, 通过 RTPCR 技术分别从叶片和果实材料中克隆获得了两条不同的 Actin 因片段生物信息学分析表明, 叶片中克隆的片段长度 604 bp, 编码 201 个氨 酸;果实中克隆的片段长度 677 bp, 编 码 225 个氨 酸两条序列与其它物种 Actin 因的碱 序列相似度高于 80% , 氨 酸序列相似度高于 93% 将来自叶片和 果实的两条序列分别命名为 VsACT7和 VsACT11, 并在 Gen ank注册, 登录号分别为 HM004434和 GU946218 : 箭筈豌豆 Actin基因 RTPCR 同源克隆 Cloning and Sequencing of Two Actin Genes from Vicia sativa Zhang Lei Liu Zhipeng Zhang Jiyu ZhangM iaoqing W angYanrong (School of PastoralAgriculture Science and Technology, Lanzhou University, Lanzhou 730000) Abstrac:t To provide internal standard for expression analysis of other genes, a partial sequence of actin gene w as isolated from common vetch ( Vicia sativa). In this study, a pair ofprmi erswas designed according to homologous actin genes among closely related plants, and then the reverse transcription PCR( RTPCR ) w as performed. Then two different sequences w ere deprived from leaf and fruit, respectively. Homological analysis show ed that the smi ilarity betw een two sequences and other actin sequencesw ere over 80% in nucleoride leve,l while 93% in anmi o acid leve.l So the two sequencesw ere considered to be different copies of actin gene in Vicia sati va. They were named asV sACT7 and VsACT11 respectively, and subm ited to Gen ank ( accession number: HM 004434, GU946218). K ey w ords: Common vetch( Vicia sativa) A ctin gene RTPCR Homocloning (A ctin) 筈豌豆( Vicia sativa)是豆科野豌豆属一年生