华北农学报 ›› 2018, Vol. 33 ›› Issue (6): 24-32. doi: 10.7668/hbnxb.2018.06.004

所属专题: 生物技术

• 论文 • 上一篇    下一篇

雪菜BjSnRK2C基因的克隆及生物信息学分析

张春兰1, 满丽莉1, 向殿军2, 李旭新2, 包乌日娜2, 刘鹏2   

  1. 1. 内蒙古民族大学 生命科学学院, 内蒙古 通辽 028042;
    2. 内蒙古民族大学 农学院, 内蒙古 通辽 028042
  • 收稿日期:2018-08-08 出版日期:2018-12-28
  • 通讯作者: 向殿军(1978-),男,黑龙江望奎人,副教授,博士,主要从事植物非生物胁迫研究;刘鹏(1973-),男,内蒙古赤峰人,教授,博士,主要从事作物遗传育种研究。
  • 作者简介:张春兰(1972-),女,内蒙古赤峰人,实验师,硕士,主要从事作物生物技术研究。
  • 基金资助:
    国家自然基金项目(31860389);内蒙古自治区自然科学基金项目(2018MS03057);内蒙古民族大学博士科研启动基金项目(BS404);内蒙古自治区饲用作物工程技术研究中心开放课题(MDK2017004;MDK2018033)

Cloning and Bioinformatics Analysis of BjSnRK2C Gene from Potherb Mustard

ZHANG Chunlan1, MAN Lili1, XIANG Dianjun2, LI Xuxin2, BAO Wurina2, LIU Peng2   

  1. 1. College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028042, China;
    2. College of Agriculture, Inner Mongolia University for Nationalities, Tongliao 028042, China
  • Received:2018-08-08 Published:2018-12-28

摘要: SnRK2家族基因编码植物体内一类丝氨酸/苏氨酸蛋白激酶,是ABA通路中影响植物抗逆能力的关键调控基因。基于SnRK2成员cDNA的保守区序列信息,利用RT-PCR与SON-PCR相结合的方法从雪菜中克隆了一个SnRK2全长cDNA序列,命名为BjSnRK2C。测序结果显示,该cDNA序列全长1 380 bp,包含一个137 bp的3'-非翻译区,一个214 bp的5'-非翻译区,一个1 029 bp的完整开放阅读框,编码342个氨基酸,在GenBank数据库的登录号为MF983711。BjSnRK2C蛋白激酶理论分子量为38.2 ku,理论等电点为5.61,属于亲水性蛋白,存在2个显著跨膜结构域,含有30处磷酸化位点,没有信号肽,最可能定位的位置是在细胞质上。BjSnRK2C蛋白激酶的二级结构具有130个α-螺旋、126个无规则卷曲、59个延伸链和27个β-转角。BjSnRK2C蛋白包含1个丝氨酸/苏氨酸酶活性结构域和1个ATP结合位点,同拟南芥AtSnRK2.8蛋白激酶亲缘关系最近,处在同一进化分枝。雪菜BjSnRK2C蛋白激酶的三级结构与拟南芥AtSnRK2.8蛋白激酶极为相似,推测它们具有类似的功能。BjSnRK2C蛋白激酶的C末端仅含有结构域Ⅰ,缺少结构域Ⅱ,暗示雪菜BjSnRK2C基因在参与非生物胁迫过程中很可能也是不依赖ABA调控通路的。研究结果可为今后深入研究BjSnRK2C功能奠定理论基础。

关键词: 雪菜, BjSnRK2C基因, 克隆, 生物信息学

Abstract: The SnRK2 family genes, which are key regulators of plant resistance to abiotic stresses in the ABA pathway, encode a class of serine/threonine protein kinases in plants. Based on the conservative cDNA sequences of SnRK2 members, a full-length cDNA sequence of SnRK2, named as BjSnRK2C, was cloned from Brassica juncea by RT-PCR and SON-PCR methods. The sequencing results showed that the length of full-length cDNA (GenBank accession No. MF983711)was 1 380 bp containing a 3'-UTR of 137 bp, a 5'-UTR of 214 bp, and a complete open reading frame of 1 029 bp with 342 amino acids. The putative molecular weight and theory isoelectric point of the BjSnRK2C protein were 38.2 ku and 5.61, respectively. The BjSnRK2C was a hydrophilic protein with 30 phosphorylation sites, contained two significant transmembrane domains, contained no signal peptide, and might played a physiological role in the cytoplasm. The secondary structure of the BjSnRK2C protein contained 130 α-helixes, 126 random coils, 59 extended strands, and 27 β-turns. The BjSnRK2C protein contained a serine/threonine enzyme activity domain and an ATP binding site, and was at the same evolutionary branch with AtSnRK2.8 from Arabidopsis. The three-dimensional structure of the BjSnRK2C protein kinase was very similar to that of AtSnRK2.8 protein kinase from Arabidopsis, suggesting that they had similar functions. The C terminal of the BjSnRK2C protein kinase contained only the domain Ⅰ, and lacked the domain Ⅱ, suggesting that the BjSnRK2C gene was probably independent of ABA regulatory pathway in abiotic stress process, which will lay a theoretical foundation for further studying the BjSnRK2C gene function.

Key words: Brassica juncea, BjSnRK2C gene, Cloning, Bioinformatics

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引用本文

张春兰, 满丽莉, 向殿军, 李旭新, 包乌日娜, 刘鹏. 雪菜BjSnRK2C基因的克隆及生物信息学分析[J]. 华北农学报, 2018, 33(6): 24-32. doi: 10.7668/hbnxb.2018.06.004.

ZHANG Chunlan, MAN Lili, XIANG Dianjun, LI Xuxin, BAO Wurina, LIU Peng. Cloning and Bioinformatics Analysis of BjSnRK2C Gene from Potherb Mustard[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(6): 24-32. doi: 10.7668/hbnxb.2018.06.004.

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