›› 2010, Vol. 18 ›› Issue (5): 683-688.DOI: 10.11733/j.issn.1007-0435.2010.05.013

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Cloning and sequence analysis of K+ channel PtAKT1 gene fragment from halophyte (Puccinellia tenuiflora)

GUO Qiang, ZHOU Xiang-rui, WANG Pei, ZHANG Jin-lin, BAO Ai-ke, WU Guo-qiang, WANG Suo-min   

  1. School of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, Gansu Province 730000, China
  • Received:2010-05-20 Revised:2010-09-17 Online:2010-10-15 Published:2010-10-15

盐生植物小花碱茅K+通道PtAKT1基因片段的克隆及序列分析

郭强, 周向睿, 王沛, 张金林, 包爱科, 伍国强, 王锁民   

  1. 兰州大学草地农业科技学院, 甘肃, 兰州, 730020
  • 通讯作者: 王锁民,E-mail:smwang@lzu.edu.cn
  • 作者简介:郭强(1981- ),男,甘肃天水人,博士研究生,研究方向为植物逆境生理与分子生物学,E-mail:guoyanbai@163.com
  • 基金资助:
    国家973项目(2007CB108901);863计划项目(2006AA10Z126);国家自然科学基金(30770347)(30700562)资助

Abstract: Degenerate primers were designed based on the conserved sequences of the AKT1 genes from other plants for investigating molecular mechanisms of K+/Na+ selective absorption in P.tenuiflora.Total RNA was extracted from the roots of Puccinellia tenuiflora to obtain an AKT1 gene fragment by reverse transcription polymerase chain reaction(RT-PCR).The sequencing result shows that the AKT1 gene fragment(about 1019 bp) from Puccinellia tenuiflora encodes 339 amino acids.Homology comparison show that the AKT1 gene fragment contains over 85% nucleotide and 73% amino acid homology when compared with other plants in the GenBank having an AKT1 gene.These results offer further understanding for full-length AKT1 gene cloning,gene expression regulation and crop improvement.

Key words: Puccinellia tenuiflora, AKT1 gene, Cloning, Sequence analysis

摘要: 为研究小花碱茅(Puainellia tenuiflora)K+/Na+选择吸收分子机制,根据已报道的其他植物AKT1基因的保守序列设计一对简并性引物,以小花碱茅根部总RNA为模板,采用RT-PCR方法克隆出AKT1基因,以期为小花碱茅AKT1全长基因的克隆、表达调控及其作物改良的研究奠定基础。序列分析结果表明:该基因长度大约为1019 bp,编码339个氨基酸;所得序列与GenBank中注册的高等植物AKT1基因序列的同源性均在85%以上,与其他AKT1的氨基酸序列同源性达73%以上。

关键词: 小花碱茅, AKT1基因, 克隆, 序列分析

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