华北农学报 ›› 2017, Vol. 32 ›› Issue (6): 8-13. doi: 10.7668/hbnxb.2017.06.002

所属专题: 油料作物 抗旱节水 生物技术

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大豆sbp基因表达方式分析及其启动子调控活性研究

赵艳, 翟莹, 李珊珊, 杨晓杰, 刘丽丽   

  1. 齐齐哈尔大学 生命科学与农林学院, 黑龙江 齐齐哈尔 161006
  • 收稿日期:2017-09-10 出版日期:2017-12-28
  • 作者简介:赵艳(1981-),女,吉林人,副教授,博士,主要从事植物分子生物学研究。
  • 基金资助:
    黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2016090)

The Study of the Expression Pattern of Soybean sbp Gene and Activity Analysis of It's Promoter

ZHAO Yan, ZHAI Ying, LI Shanshan, YANG Xiaojie, LIU Lili   

  1. College of Life Science and Agroforestry, Qiqihaer University, Qiqihaer 161006, China
  • Received:2017-09-10 Published:2017-12-28

摘要: 研究大豆蔗糖结合蛋白基因(sbp)的表达方式及其启动子的调控活性,为揭示sbp基因表达本质及其启动子功能研究、利用提供理论依据。利用qRT-PCR方法,检测sbp基因在不同逆境胁迫条件下及不同组织中的表达方式;利用PCR方法,克隆sbp基因5'端上游序列并对其进行预测分析;在转基因烟草中,研究sbp基因启动子在干旱胁迫条件下及不同组织中的调控活性。sbp基因受干旱诱导上调,而在盐、低温、ABA诱导下表达下降。sbp基因在大豆根、茎、叶、花中的相对表达量低,而在种子中的相对表达量高。克隆获得sbp基因5'端上游942 bp序列,命名为SP,预测分析表明,SP序列中含有多种典型的种子特异表达元件及与激素、逆境诱导相关的元件。组织化学分析表明,在转基因烟草中,SP启动子驱动gus基因在干旱胁迫下及种子中高表达。推测SP启动子兼具干旱诱导表达活性和种子特异表达特性。

关键词: 大豆, sbp基因, 启动子, 克隆, 转基因烟草

Abstract: The expression pattern of soybean sbp gene and activity analysis of it's promoter was studied,which provided the theoretical foundation for further understanding the regulation of sbp gene expression and utilizing the promoter. The expression pattern of soybean sbp gene under different stresses and in soybean tissues were detected by qRT-PCR. The 5'-flanking upstream sequence of soybean sbp gene was isolated by PCR method and analyzed by some prediction softwares. The activity analysis of the promoter under drought stress and in different tissues were performed in transgenic tabacco. Soybean sbp gene was up-regulated by drought and were down-regulated by salt,cold and ABA. There was a little activity in roots,stems,leaves and flowers,but there was highest activity in seeds. The 5'-flanking upstream sequence of soybean sbp gene,named SP,was isolated,and the length was 942 bp. Sequence analysis revealed that this fragment contained a series of motifs related to seed-specific promoters,and stress-inducible promoters. The gus gene driven by SP promoter had high expression activity under drought stress or/and in seeds in transgenic tabacco through GUS histochemical analysis. SP promoter could have characteristics of drought-induced and seed-specific activity.

Key words: Soybean, sbp gene, Promoter, Cloning, Transgenic tobacco

中图分类号: 

引用本文

赵艳, 翟莹, 李珊珊, 杨晓杰, 刘丽丽. 大豆sbp基因表达方式分析及其启动子调控活性研究[J]. 华北农学报, 2017, 32(6): 8-13. doi: 10.7668/hbnxb.2017.06.002.

ZHAO Yan, ZHAI Ying, LI Shanshan, YANG Xiaojie, LIU Lili. The Study of the Expression Pattern of Soybean sbp Gene and Activity Analysis of It's Promoter[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 8-13. doi: 10.7668/hbnxb.2017.06.002.