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当前位置: 首页 > 文章 > 玉米ZmCIPK6基因的克隆及植物表达载体的构建 湖北农业科学 2015,54 (21) 5432-5435
Position: Home > Articles > Cloning of ZmCIPK6 Gene and Construction of Its Plant Expression Vector Hubei Agricultural Sciences 2015,54 (21) 5432-5435

玉米ZmCIPK6基因的克隆及植物表达载体的构建

作  者：
李横江;黄慧艳;张小菊
单  位：
华中科技大学武昌分校城市建设学院
关键词：
玉米;ZmCIPK6基因;植物表达载体
摘  要：
根据MaizeDGB数据库中ZmCIPK6基因序列设计引物,采用RT-PCR技术从玉米中克隆了1个ZmCIPK6基因。ZmCIPK6基因c DNA全长1 739 bp,5′-非编码区长281 bp,3′-非编码区长111 bp,编码区长1 347 bp,编码448个氨基酸,预测分子量为48.8 KDa,等电点为9.14。氨基酸同源性分析发现,ZmCIPK6与高粱SbCIPK6同源性较高。根据ZmCIPK6的基因序列和植物表达载体pCAMBIA3301的多克隆位点设计带有限制性内切酶位点的特异性引物,以质粒pMD18-T-ZmCIPK6为模板,PCR扩增ZmCIPK6基因片段,双酶切目的片段及载体,回收后连接,构建成该基因的植物表达载体。经PCR检测及测序鉴定,表明成功构建了ZmCIPK6基因的植物表达载体,为进一步遗传转化研究基因的功能奠定基础。
译  名：
Cloning of ZmCIPK6 Gene and Construction of Its Plant Expression Vector
作  者：
LI Heng-jiang;HUANG Hui-yan;ZHANG Xiao-ju;Urban Construction School, Huazhong University of Science and Technology Wuchang Branch;
关键词：
maize;;ZmCIPK6 gene;;plant expression vector
摘  要：
According to the sequence of ZmCIPK6 in Maize DGB database,primers were designed. RT-PCR method was used to clone ZmCIPK6 gene. A gene coding for ZmCIPK6 was isolated from maize(Zea mays).The full length ZmCIPK6 c DNA was1 739 bp,including a 281 bp 5′-UTR,an ORF of 1 347 bp,and a 111 bp 3′-UTR.This c DNA sequence encoded a polypepide of 448 amino acid residues with a predicted molecular mass of 48.8 k Da and a basic isoelectric point of 9.14. The deduced amino acid sequence had a high homology with Sb CIPK6 from Sorghum bicolor.According to the restriction enzyme sites of expression vector pCAMBIA3301 and sequence of ZmCIPK6 gene,one pair of primers containing restriction enzyme sites were designed. The ZmCIPK6 gene was obtained from pMD18-T-ZmCIPK6 by PCR. PCR product and the plasmid pCAMBIA3301 were digested by the corresponding restricted enzymes respectively,and then the ZmCIPK6 gene was cloned into pCAMBIA3301 vector.PCR and sequencing results suggest that plant expression vector of ZmCIPK6 gene was constructed successfully.

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