生物技术通报 ›› 2013, Vol. 0 ›› Issue (4): 55-62.

• 研究报告 • 上一篇    下一篇

盐芥EhKCR1 基因的克隆及表达分析

徐小静 安会灵 韦百阳 周宜君 冯金朝   

  1. 中央民族大学生命与环境科学学院,北京 100081
  • 收稿日期:2012-12-25 修回日期:2013-04-22 出版日期:2013-04-22 发布日期:2013-04-22
  • 作者简介:徐小静,女,博士,副教授,研究方向:植物分子生物学及植物保护学;E-mail: xuxiaojing@ muc.edu.cn
  • 基金资助:
    国家自然科学基金项目(31100287),中央高校基本科研业务费专项资金(1112KYZY43),中央民族大学“985”项目(MUC98504-14, MUC98507-08)

Real-time PCR Cloning and Expression of EhKCR1 from Eutrema halophilum

Xu Xiaojing, An Huiling, Wei Baiyang, Zhou Yijun, Feng Jinchao   

  1. College of Life and Environmental Sciences,Minzu University of China,Beijing 100081
  • Received:2012-12-25 Revised:2013-04-22 Published:2013-04-22 Online:2013-04-22

摘要: 以山东生态型盐芥(Eutrema halophilum)为试材,采用电子克隆及RT-PCR 的方法,从中获得一个β-酮脂酰-CoA 还原酶基因全长cDNA,命名为EhKCR1,GenBank 登录号为JQ389855。EhKCR1 cDNA 长1 152 bp,含954 bp 开放阅读框架,编 码318 个氨基酸。采用生物信息学的手段对EhKCR1 蛋白的保守结构域、疏水性和二级结构等进行了分析。EhKCR1 蛋白具有 NADH 结合结构域[G(X)3GXG(X)3A(X)3A(X)2G]负责裂解的关键结构域[Y(X)3K]。序列比对和系统进化分析表明, 盐芥EhKCR1 基因与来源于拟南芥和油菜的KCR1 亲缘关系最近,与单子叶植物来源的KCR1 亲缘关系较远。Real-time PCR 分析 表明EhKCR1 受ABA 和干旱明显诱导。推测EhKCR1 与盐芥的抗逆有关。

关键词: 盐芥, EhKCR1, 电子克隆, 生物信息学分析

Abstract: A cDNA was cloned from Eutrema halophilum(Shangdong ecotype)using in silico cloning and RT-PCR methods. This gene was predict coding β-ketoacyl-CoA reductase, named as EhKCR1, with GenBank accession number JQ389855. The full length is 1 152 bp and contains a complete ORF with 954 bp encoding 318 amino acid. The conservative domain, hydrophobicity and second structure of EhKC1 protein were predicted by bioinformatic analysis. EhKCR1 has the putative NADH binding motif[G(X)3GXG(X)3A(X)3A(X)2G]and the essential catalytic motif(Y(X)3K). Alignment of predicted amino acid sequence of EhKCR1 with other plants and phylogenetic analysis of various EhKCR1 homologues found that EhKCR1 was very close to that from Arabidopsis thaliana and Brassica napus, but far from to EhKCR1 of monocotyledon. The results of Real-time PCR indicated that EhKCR1 was induced by ABA and drought. Therefore EhKCR1 may be connected with the strong stress tolerence of Eutrema halophilum.

Key words: Eutrema halophilum, EmKCR1 In silico cloning, Bioinformatic analysis, Real-time PCR