大豆GmNHX3基因克隆及遗传转化载体的构建

J4 ›› 2012, Vol. 50 ›› Issue (02): 365-370.

大豆GmNHX3基因克隆及遗传转化载体的构建

张继星^1,2, 王晓宇³, 陈永胜^1,2, 黄凤兰^1,2, 李国瑞^1,2

1. 内蒙古民族大学生命科学学院, 内蒙古通辽 028000；2. 内蒙古自治区高校蓖麻产业工程技术研究中心, 内蒙古通辽 028000；3. 吉林大学
植物科学学院, 长春 130062

收稿日期:2011-11-27 出版日期:2012-03-26 发布日期:2012-03-21
通讯作者: 张继星 E-mail:jxzhang@imun.edu.cn

Cloning of GmNHX3 Gene from Glycine max L Merr andConstruction of Its Genetic Transformation Vector

ZHANG Jixing^1,2, WANG Xiaoyu³, CHEN Yongsheng^1,2, HUANG Fenglan^1,2, LI Guorui^1,2

1. College of Life Sciences, Inner Mongolia University for the Nationalities, Tongliao 028000, Inner Mongolia AutonomousRegion, China|2. Engineering Research Center of Castor of Universities of Inner Mongolia Autonomous, Tongliao 028000,Inner Mongolia Autonomous Region, China|3. College of Plant Sciences, Jilin University, Changchun 130062, China

Received:2011-11-27 Online:2012-03-26 Published:2012-03-21
Contact: ZHANG Jixing E-mail:jxzhang@imun.edu.cn

摘要/Abstract

摘要：

利用RACE(rapidamplification of cDNA end)方法, 以大豆叶片提取总的RNA为模板克隆了大豆Na⁺/H⁺逆向转运蛋白（Glycine max Na⁺/H⁺ antiporter, GmNHX）基因, 并将其连接到表达载体PBI121中, 构建重组表达载体PBI121NHX3. 分析结果表明：该基因的ORF为1 503 bp, 推测编码501个氨基酸. 与所选取的10种植物同类蛋白氨基酸序列进行对比, 一致性为72%～94%, 并具有真核生物单价阳离子(氢离子)反向转运蛋白典型的结构域, 将该基因命名为GmNHX3, GenBank接收号为JN872904. 通过PCR和酶切鉴定, PBI121NHX3构建成功.

关键词: 大豆； Na⁺/H⁺逆向转运蛋白；载体构建

Abstract:

RACE (rapidamplification of cDNAends) technique was used to clone Na⁺/H⁺ antiporter gene from Glycine max L Merr with the total RNA as the template. This gene was linked to the plasmid PBI121 and the expression vector PBI121NHX3 was constructed. The results show its ORF（open reading frame）consists of 1 503 bp, which can be deduced to encode 501 amino acids. It was found that predicted amino acid sequence had an identification of 72%— 94% compared with amino acid sequence from other ten plant species and it had a typical structural domain of the monovalent cation(proton) antiporter. The gene was named GmNHX3, (GenBank accession number: JN872904). Genetic transformation vector of PBI121NHX3 was constructed successfully by identification method of PCR and restriction enzyme digestion.

Key words: Glycine max L Merr, Na⁺/H⁺ antiporter; construction vector

中图分类号:

Q785

张继星, 王晓宇, 陈永胜, 黄凤兰, 李国瑞. 大豆GmNHX3基因克隆及遗传转化载体的构建[J]. J4, 2012, 50(02): 365-370.

ZHANG Ji-Xing, WANG Xiao-Yu, CHEN Yong-Qing, HUANG Feng-Lan, LI Guo-Rui-. Cloning of GmNHX3 Gene from Glycine max L Merr andConstruction of Its Genetic Transformation Vector[J]. J4, 2012, 50(02): 365-370.