Expression Analysis of a Stress Repressed Gene OsDSR4 from DUF966 Family and Generation of OsDSR4- overexpressing Transgenic Rice(Oryza sativa L. ssp. japonica)
OSdSr4基因是duf966基因家族中的一个未知功能基因,目前其生物学功能尚不清楚。本研究生物信息学分析显示,OSdSr4基因CdnA全长2 167 bP,包含一个1 149 bP的开放阅读框(Orf),编码382个氨基酸,推测的蛋白中包含一个高度保守的duf966结构域;表达模式分析表明,OSdSr4主要在水稻(OryzA SATIVA l.SSP.JAPOnICA)的茎节间和叶片中表达,干旱、高盐和低温等非生物胁迫明显抑制了OSd Sr4的表达,而脱落酸(AbSCISIC ACId,AbA)则显著诱导了它的表达;利用重叠延伸PCr方法成功克隆了OSdSr4,并将其转化进水稻中,获得了32株超表达转基因植株。分子鉴定结果表明,该基因已被整合进水稻基因组中,并在部分转基因植株中实现了超量表达。本实验为进一步开展OSdSr4基因的生物学功能研究提供了基础资料。OsDSR4 is a gene of unkown function in DUF966 gene family,and the function of DUF966 family genes have not been reported until now.In this study,the bioinformatic analysis showed that the cDNA of OsDSR4 had 2 167 bp containing an open reading frame(ORF) of 1 149 bp,and it encoded a putative protein of 372 amino acids with a highly conserved DUF966 domain.The gene expression profile analysis indicated that OsDSR4 was expressed mainly in internode and leaf blade of rice(Oryza sativa L.),and it was repressed markedly by drought,salt and cold stresses,and induced significantly by abscisic acid(ABA).OsDSR4 was cloned using overlap extension PCR,and the fusion construct containing OsDSR4 was introduced into rice(Oryza sativa L.ssp.japonica) by Agrobacterium- mediated transformation method.Thirty- two OsDSR4- overexpressing transgenic plants were obtained and identified by PCR and qRT-PCR,which was demonstated that OsDSR4 had been integrated into rice genome and was overexpressed in some positive transgenic plants.These results establish the foundation for further study of the precise function of OsDSR4.国家重点基础研究发展计划(973)前期研究专项(No.2012CB126312
