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水稻OsMAPK17-1基因的分离及转基因研究
引用本文:肖文芳,陈艳,汪德锋,符秀梅,肖晓蓉,黎秀琼,庞金环,陈银华. 水稻OsMAPK17-1基因的分离及转基因研究[J]. 热带作物学报, 2013, 34(8): 1490-1497
作者姓名:肖文芳  陈艳  汪德锋  符秀梅  肖晓蓉  黎秀琼  庞金环  陈银华
作者单位:1 海南大学热带生物资源可持续利用重点实验室 3 海南大学环境与植物保护学院;中国科学院微生物研究所;海南大学热带生物资源可持续利用重点实验室;海南大学热带生物资源可持续利用重点实验室;海南大学热带生物资源可持续利用重点实验室;中国科学院微生物研究所;中国科学院微生物研究所;海南大学热带生物资源可持续利用重点实验室
基金项目:转基因生物新品种培育重大专项“水稻广谱抗病基因的系统筛选及新种质创制”(No. 2009ZX08009-41B);国家自然基金(No.31260345);海南省重大科技项目(No. 2D2X2013023-2)。
摘    要:OsMAPK17-1作为一种MAP激酶,受多种生物和非生物胁迫的诱导,且对病原菌的侵染有一定的防卫作用。为研究水稻OsMAPK17-1基因的功能,通过RT-PCR方法分离OsMAPK17-1基因全长cDNA,分别构建过表达与RNAi植物表达载体,采用农杆菌介导法以粳稻日本晴为受体进行遗传转化,PCR检测具有潮霉素抗性基因的转基因水稻植株,结果表明:OsMAPK17-1基因及RNAi片段均已整合到水稻基因组中。利用PCR和RT-PCR的方法对转基因后代进行筛选,获得过表达和RNAi转基因纯系各1个,模拟干旱处理结果表明,RNAi株系在干旱处理下比对照更有利于根的生长,这些结果为进一步研究OsMAPK17-1基因的功能奠定了基础。

关 键 词:水稻  OsMAPK17-1基因  遗传转化  过表达载体  RNAi载体

Isolation of Rice OsMAPK17-1 Gene and Its Genetic Transformation
XIAO Wenfang,CHEN Yan,WANG Defeng,FU Xiumei,XIAO Xiaorong,LI Xiuqiong,PANG Jinhuan and CHEN Yinhua. Isolation of Rice OsMAPK17-1 Gene and Its Genetic Transformation[J]. Chinese Journal of Tropical Crops, 2013, 34(8): 1490-1497
Authors:XIAO Wenfang  CHEN Yan  WANG Defeng  FU Xiumei  XIAO Xiaorong  LI Xiuqiong  PANG Jinhuan  CHEN Yinhua
Affiliation:1 Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University 3 College of Environment and Plant Protection, Hainan University;Institute of Microbiology, Chinese Academy of Science;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University;Institute of Microbiology, Chinese Academy of Science;Institute of Microbiology, Chinese Academy of Science;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, Hainan University
Abstract:OsMAPK17-1, a MAP Kinase in rice, is induced by various biotic and abiotic stress and involved in the regulation of plant defense responses. To research the functions of rice gene OsMAPK17-1, the full length cDNA of OsMAPK17-1 was isolated by RT-PCR amplification, the over-expression and RNA interference vectors were constructed with modified pCAMBIA1300 and pTCK303 as framework plasmid, respectively, with the Hygromycin phosphotransferase gene as a selectable marker. Then the vectors were respectively transferred into the rice through Agrobacterium-mediated transformation. PCR analysis confirmed that OsMAPK17-1 and its RNAi were integrated into rice genome. The homologous transgenic lines were screened with the method of PCR. The RNAi lines were more suitable for the drought stress. These works laid the foundation for further study of OsMAPK17-1.
Keywords:
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