水稻OsCERK2转基因植株的获得及初步分析  

Obtaining and the Preliminary Analysis of Rice OsCERK2 Transgenic Plants

作　　者：汪德锋[1,2] 陈艳[3] 肖文芳[1,2] 符秀梅[1,2] 肖晓蓉[1,2] 黎秀琼[3] 庞金环[3] 陈银华[1,2] 

机构地区：[1]海南大学海南省热带生物资源可持续利用重点实验室,海南海口570228 [2]海南大学农学院,海南海口570228 [3]中国科学院微生物研究所,北京10010

出　　处：《热带生物学报》2013年第2期146-152,159,共8页Journal of Tropical Biology

基　　金：国家自然科学基金项目(31260345);转基因生物新品种培育重大专项(2009ZX08009-41B)

摘　　要：以水稻粳稻品种日本晴为研究材料,利用拟南芥CERK1的蛋白质序列检索,在水稻基因组候选了与拟南芥CERK1同源的水稻基因OsCERK2,通过RT-PCR分离了该基因的全长cDNA。生物信息学分析显示,OsCERK2是一种含有信号肽的质膜蛋白,胞外结构域含有LsyM基序,激酶结构域含有酪氨酸蛋白激酶结构域。构建了由35S启动子驱动该基因的过表达遗传转化载体和由玉米的泛素基因的启动子驱动的RNA干涉(RNAi)的遗传转化载体,利用农杆菌介导的遗传转化技术,将OsCERK2基因导入水稻,得到T0代转基因植株。对T0代植株进行了PCR检测和半定量RT-PCR检测,获得了OsCERK2有效表达的转基因植株。In this study,Oryza sativa ssp. japonica var. Nipponbare used as material,the Arabidopsis CERK1 was used to retrieve its homologous genes in the rice genome and a rice homologous gene CERK2,OsCERK2 was obtained,which cDNA of OsCERK2 was isolated by RT-PCR. Bioinformatics analysis showed that OsCERK2 was a plasma membrane protein including a signal peptide,its extracellular domain included an LsyM domain,its kinase domain included a protein tyrosine kinase domain. Overexpression vector driven by 35S promoter and RNA interference （ RNAi） vector driven by maize ubiquitin promoter were constructed. The OsCERK2 gene was then transferred into rice by Agrobacterium-mediated transformation. T 0 transgenic plants were obtained from each vector. Amplifying T 0 transgenic plants by PCR and RT-PCR showed transgenic plants with efficient expression were gained. The results would lay a foundation on further studying functions of OsCERK2.

关 键 词：水稻(Oryza sativa) OsCERK2 基因分离 载体构建 遗传转化 RT-PCR 

分 类 号：Q812[生物学—生物工程]