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[目的]克隆线粒体相关基因nad1,获得转nad1的转基因水稻植株。[方法]采用TRIzol法提取水稻幼苗总RNA,以反转录的cDNA为模板,扩增得到nad1;将nad1接到线粒体信号肽Rf1b的5 (Rf1b5 ),装载到pCAMBIA1305.1双元表达载体,采用农杆菌介导的愈伤组织侵染法进行遗传转化。[结果]克隆的目的基因nad1大小为978 bp,成功构建了携带线粒体信号肽的nad1植物表达载体,并获得了转nad1基因的阳性植株。[结论]为探讨水稻中过表达nad1对水稻生长的影响奠定了基础。
[Objective] To clone mitochondria-related gene nad1 and obtain transgenic rice plants transformed with nad1. [Method] TRIzol method was used to extract total RNA of rice seedlings and nad1 was amplified by reverse transcribed cDNA. The nad1 was inserted into Rf1b5 of mitochondrial signal peptide Rf1b5, and then was inserted into pCAMBIA1305.1 binary vector. Agrobacterium tumefaciens-mediated callus infection was used for genetic transformation. [Result] The nad1 gene of cloned gene was 978 bp in length. The nad1 plant expression vector carrying the mitochondrial signal peptide was successfully constructed and the transgenic plant with nad1 gene was obtained. [Conclusion] The study laid the foundation for exploring the effect of nad1 overexpression in rice on rice growth.