中国农业科学 ›› 2009, Vol. 42 ›› Issue (10): 3600-3608 .doi: 10.3864/j.issn.0578-1752.2009.10.0027

• 园艺 • 上一篇    下一篇

普通烟草CDPK基因家族的克隆及表达分析

太帅帅,刘贯山,孙玉合,陈珈

  

  1. (中国农业科学院烟草研究所/中国农业科学院烟草遗传改良与生物技术重点开放实验室)

  • 收稿日期:2009-01-06 修回日期:2009-02-24 出版日期:2009-10-10 发布日期:2009-10-10
  • 通讯作者: 孙玉合,刘贯山

Cloning and Expression of Calcium-Dependent Protein Kinase (CDPK) Gene Family in Nicotiana tabacum

TAI Shuai-shuai, LIU Guan-shan, SUN Yu-he, CHEN Jia
  

  1. (中国农业科学院烟草研究所/中国农业科学院烟草遗传改良与生物技术重点开放实验室)

  • Received:2009-01-06 Revised:2009-02-24 Online:2009-10-10 Published:2009-10-10
  • Contact: SUN Yu-he, LIU Guan-shan

摘要:

【目的】从烟草中分离更多CDPK基因,分析该基因家族的序列特征、进化关系和表达谱,为全面揭示其功能奠定基础。【方法】采用RT-PCR、RACE和生物信息学的方法,分离普通烟草CDPK基因;采用MEGA4.0软件构建系统进化树;利用RT-PCR研究普通烟草全长基因的表达谱。【结果】分离到普通烟草CDPK基因8个,其中3个可以找到完整的开放读码框;系统进化树分析显示,烟草属11个全长CDPK基因位于4个亚家族,预测的2对普通烟草和拟南芥的直系同源基因在功能上可能非常保守;3个普通烟草全长基因虽然在所有的组织中都能检测到,但是它们的表达谱在不同的组织间存在明显差异。【结论】分离到8个未报道的普通烟草CDPK基因,结合已有的报道,目前在普通烟草中获得了15个CDPK基因,为在基因组范围内研究普通烟草CDPK基因家族的功能奠定了基础。

关键词: 普通烟草, CDPK, 进化树, 基因表达

Abstract:

【Objective】To further study the function of calcium-dependent protein kinase (CDPK) gene family in tobacco (Nicotiana tabacum), it is necessary to isolate CDPKs from tobacco and describe the sequence characteristics, evolutionary relationship and gene expression. 【Method】 RT-PCR, RACE and bioinformatics methods were used to isolate CDPKs from tobacco. A phylogenetic tree was created using the MEGA4.0 program and the expression patterns of the 3 full-length CDPK genes were studied by RT-PCR. 【Result】 Eight additional tobacco CDPK genes were obtained, of which 3 possessed complete ORFs. Phylogenetic analysis divided 11 full-length Nicotiana CDPK genes into 4 subfamilies, and 2 putative tobacco and Arabidopsis orthologous CDPK genes might correspond to well-conserved functions. Three full-length tobacco CDPK genes were detected in all tobacco organs tested, but the expression patterns of them were significantly different. 【Conclusion】 Eight non-redundant tobacco CDPK genes were isolated in this study. Along with the previously characterized CDPK genes, at least 15 members of the CDPK family are existing in tobacco. This work established a foundation for a genome-wide study of this important gene family in tobacco.

Key words: Nicotiana tabacum, CDPK, phylogenetic tree, gene expression