普通烟草CDPK基因家族的克隆及表达分析  

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

作　　者：太帅帅[1] 刘贯山[1] 孙玉合[1] 陈珈[2] 

机构地区：[1]中国农业科学院烟草研究所/农业部烟草类作物质量控制重点开放实验室,山东青岛266101 [2]中国农业大学生物学院/植物生理学与生物化学国家重点实验室,北京100193

出　　处：《中国农业科学》2009年第10期3600-3608,共9页Scientia Agricultura Sinica

基　　金：国家烟草专卖局重点资助项目(110200701021;110200701022);植物生理学与生物化学国家重点实验室开放课题基金(PPB08004)

摘　　要：【目的】从普通烟草中分离更多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。【Objective】To further study the function of calcium-dependent protein kinase （CDPK） gene family in common tobacco （Nicotiana tabacum）,it is necessary to isolate more CDPKs from common tobacco and describe the sequence characteristics,evolutionary relationship and gene expression. 【Method】 RT-PCR,RACE and bioinformatics methods were used to isolate CDPKs from common 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 common 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 common tobacco and Arabidopsis orthologous CDPK genes might correspond to well-conserved functions. Three full-length common tobacco CDPK genes were detected in all common tobacco organs tested,but their expression patterns of them were significantly different. 【Conclusion】 Eight non-redundant common tobacco CDPK genes were isolated in this study. Along with the previously characterized CDPK genes,at least 15 members of the CDPK family exist in common tobacco. This work established a foundation for a genome-wide study of this important gene family in common tobacco.

关 键 词：普通烟草 CDPK 进化树 基因表达 

分 类 号：S572[农业科学—烟草工业]