水稻逆境响应基因OsMsr15的表达分析与克隆

应用Affymetrix水稻表达芯片,分析了苗期和孕穗期的培矮64S水稻基因在高温、干旱和低温逆境处理条件下的表达模式,发现多个与逆境反应相关基因.其中,OsMsr15(Oryza sativa L.multiple stresses responsivegene 15)受高温、干旱和低温胁迫诱导,表达水平均显著上调.实时定量PCR分析结果与芯片数据基本吻合.序列分析表明,该基因无内含子,ORF(open reading frame)全长为717 bp,编码一个由238个氨基酸残基组成、具有典型C2H2结构域的锌指蛋白,推测相对分子质量约为2.46 kD,pI值约为8.90,蛋白分子的氨基端和羧基端分别存在一个推测的核定位信号B-box和EAR(ERF-associated amphiphilic repression)基序.对OsMsr15可能的启动子区域进行分析,发现多种与逆境诱导相关的调控元件.该基因编码的蛋白在不同物种中存在高度相似性,显示OsMsr15基因可能作为一个保守的耐逆基因,参与植物的多种耐逆反应过程.

Expression and Cloning of A Stress Responsive Gene OsMsr15 in Rice

By using the GeneChip Rice Genome Array(Affymetrix),global genome expression profiles of rice Pei’ai 64S were analyzed at seeding and booting stages under heat,drought and cold stresses.A number of stress responsive genes were identified.OsMsr15 was highly reduced in response to heat,drought and cold stresses,which was matched to the results from microarray and quantitative real-time RT-PCR analyses.Se-quence analysis showed that the ORF of OsMsr15 is 717 bp in length,encods a protein of 238 amino acids with M.W≈2.46 kD and pI≈8.90.The gene had no intron,and the encoded protein contained typical conserved C2H2 domain.A putative nuclear location signal(B-box) and an ERF-associated amphiphilic re-pression motif(EAR-motif) were found in the N-terminal and C-terminal regions of the protein,respectively.Analysis of the putative promoter region found that several regulatory elements related to stress responses.Proteins with high similarity to OsMsr15 were found among different species.Thus,OsMsr15 might be a conserved candidate gene involved in plant stress tolerance.