不同花生品种B06基因的克隆与序列分析  

Cloning and sequence analysis of B06 gene in different peanut breeds

作　　者：陈泽宇[1] 左欣欣[1] 严海燕[1] 

机构地区：[1]中南民族大学生命科学学院,湖北武汉市430074

出　　处：《广西农学报》2011年第5期25-31,共7页Journal of Guangxi Agriculture

基　　金：BZY05011花生种子DOF结合蛋白功能性基因组的研究;中南民族大学大学生科研创新基金

摘　　要：Dof(DNA binding with one finger)转录因子是一类植物特有的、进化上高度保守的基因家族,对植物多方面的生理过程均发挥调控作用[1]。根据Dof家族中的一个B06基因的cDNA保守序列设计引物,对8个品种花生的B06基因进行PCR扩增克隆相似基因,并测定序列。实验成功克隆出四粒红和073103的完整B06基因和其余6个品种的大部分B06基因,其中073103含有一个内含子,其余7个品种不含内含子。通过比较8个品种间B06基因的序列差异与品种性状间的关系,并通过生物信息学对基因及编码蛋白进行分析,探究B06蛋白可能的功能。结果发现:典型的B06蛋白由176个氨基酸组成,在N-端附近37个氨基酸区域处形成保守的单锌指DOF结构;推测在DOF区域后的59-67氨基酸处有核定位信号序列;蛋白序列中有多个N-糖基化、蛋白激酶C磷酸化、酪蛋白激酶Ⅱ磷酸化、N-十四烷酰基化位点,可以接受不同途径的信号刺激,修饰前后发生定位或活性的变,推测Bo6蛋白可能与植物对外界刺激的反应相关。结合性状分析,推测B06蛋白与花生种子中的脂肪和蛋白质含量高低无明显联系。试验为进一步深入研究B06蛋白的功能奠定了基础。Dof（DNA binding with one finger）transcription factors is a highly conserved gene family, which play a role of regulation and control in various physiological processes of plant. According to conserved sequences of 1306 gene eDNA, with a pair of primers, DOF 1306 similar genes were cloned, sequenced and compared from genomic DNA of eight cultivar. The results showed that a typical B06 protein is composed of 176 animo adds, in which a DOF domain close to N-terminal with 37 animo acids forming the typical single zinc finger structure; B06 protein does not contain a signal peptide at the N-terminal, and the domain next to DOF domain with 9 animo acids are predicted as a nuclear localization signal （NLS）; there exit 3 N-glycosylation sites, 2 protein kinase C phosphorylation site, 1 casein kinase II phosphorylation site and 3 N-myristoylation sites acceptting signal stimulus via different ways, suggest that localization and activity of B06 alters before or after the modification; presumably related with the responds to the outside stimulus.

关 键 词：花生 DOF 转录因子 种子发育 蛋白质结构 

分 类 号：S565.2[农业科学—作物学]