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[目的]克隆水稻线粒体基因atp6,构建转基因表达载体,获得35S∷Rf1b5′∷atp6转基因水稻植株。[方法]设计目的基因的特异引物,采用TRIzol法提取水稻叶片总RNA,借助Toyobo反转录试剂盒反转录为cDNA,通过PCR扩增得到atp6全序列;将atp6连接到含线粒体信号肽(Rf1b5′)的pCAMBIA1302双元表达载体上,采用农杆菌介导的愈伤组织侵染法进行水稻的遗传转化[结果]利用目的基因atp6成功构建了35S∷Rf1b5′∷atp6植物表达载体,并获得了转atp6基因的阳性植株。[结论]为探讨水稻中超表达atp6对水稻生长的影响奠定基础。
[Objective] The study aimed to clone rice mitochondrial gene atp6 and construct a transgene expression vector to obtain 35S :: Rf1b5 ’:: atp6 transgenic rice plants. [Method] The specific primers of the target gene were designed and the total RNA was extracted from rice leaves by TRIzol method. The full length atp6 was amplified by PCR using Toyobo reverse transcription kit. The atp6 gene was then ligated into the plasmid containing mitochondrial signal peptide ( Rf1b5 ’) pCAMBIA1302 binary vector was used to transform rice with Agrobacterium tumefaciens-mediated callus [Result] The 35S :: Rf1b5’ :: atp6 plant expression vector was successfully constructed with the target gene atp6 and obtained Transferred top6 gene positive plants. [Conclusion] The research laid the foundation for exploring the effect of atp6 overexpression on rice growth in rice.