浙江农业学报 ›› 2017, Vol. 29 ›› Issue (3): 360-365.DOI: 10.3969/j.issn.1004-1524.2017.03.02

• 作物科学 • 上一篇    下一篇

烟草卵细胞特异表达基因的克隆与分析

罗岸   

  1. 长江大学 生命科学学院,湖北 荆州 434023
  • 收稿日期:2016-07-28 出版日期:2017-03-20 发布日期:2017-03-31
  • 作者简介:罗岸 (1984—),男,湖北荆州人,博士,讲师,主要从事植物生殖发育研究。E-mail:anluo@whu.edu.cn
  • 基金资助:
    长江大学自然科学基金项目(2014NSFY023)

Detection and analysis of an egg cell-expressed gene in Nicotiana tabacum

LUO An   

  1. College of Life Science, Yangtze University, Jingzhou 434023, China
  • Received:2016-07-28 Online:2017-03-20 Published:2017-03-31

摘要: 构建烟草卵细胞和合子的cDNA文库,通过抑制性差减杂交和镜像选择进行卵细胞特异表达基因的筛选。筛选结果得到一个卵细胞特异表达的,与胱天蛋白酶基因类似(CASP-like)的基因NtEE1。生物信息学分析表明,NtEE1蛋白由194个氨基酸组成,含有4个跨膜结构域。NtEE1-GFP融合蛋白的亚细胞定位也显示,该蛋白可能存在于质膜和核膜上。同时扩增拟南芥actin11基因的启动子序列驱动核定位GFP的表达,并对烟草进行异位转基因,结果发现,actin11在烟草卵细胞中具有较强的启动子活性。这些研究结果可以为通过RNAi和过表达来研究NtEE1基因在烟草卵细胞发育和在精、卵细胞相互作用中可能的功能提供理论基础。

关键词: 卵细胞, 跨膜结构域, 烟草, 配子相互作用

Abstract: cDNA pools of the egg cell and zygote of tobacco were constructed, and suppression subtractive hybridization and mirror orientation selection were used to select the genes specially expressed in the egg cell. So a gene named NtEE1 was found, which was CASP-like gene. NtEE1 encoded 194 amino-acid residues, in which 4 transmembrane domains existed by bioinformatics prediction. The subcellular localization of NtEE1-GFP also indicated that NtEE1 might be located on the nuclear membrane and plasma membrane. The promoter of actin11 of Arabidopsis was chosen to express nuclear-located GFP. The vector was transformed to tobacco, and the promoter activity of actin11 was proved to be intensive in the tobacco egg cell. These would provide a foothold to investigate the function of NtEE1 in the development of egg cell and the mechanism of gamete interaction in tobacco.

Key words: egg cell, transmembrane domain, tobacco, gamete interaction

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