论文部分内容阅读
本研究在GenBank中找到花粉育性恢复基因MS45、花粉致死基因ZmAA1、颜色筛选标记基因DsRed2及其特异性启动子和终止子序列,并在目的基因和植物表达载体pCAMBIA3300设计带有限制性内切酶位点的特异性引物,扩增基因片段,双酶切目的片段及载体,回收后连接,构建表达载体pCAMBIA3300-MS45-DsRed2-ZmAAI,通过农杆菌介导的萌动胚遗传转化法将构建的表达载体转化到优良玉米自交系吉A001中,通过喷洒含5mg/L除草剂筛选得到397株草胺膦抗性植株,用PCR检测得到119株含有目的基因的阳性植株。结果表明,MS45-DsRed2-ZmAAI基因在玉米中得到表达。
In this study, pollen fertility restorer MS45, pollen lethal ZmAA1, DsRed2 and their specific promoter and terminator sequences were found in GenBank. The target gene and plant expression vector pCAMBIA3300 were designed with restriction endonuclease Enzyme-site-specific primers, amplified gene fragments, double digestion of the target fragment and vector, recovered and ligated to construct the expression vector pCAMBIA3300-MS45-DsRed2-ZmAAI, Agrobacterium-mediated embryo genetic transformation of the constructed The expression vector was transformed into a good maize inbred line Ji A001, and 397 glufosinate-resistant plants were screened by spraying 5 mg / L herbicide. 119 positive plants containing the gene of interest were obtained by PCR. The results showed that the MS45-DsRed2-ZmAAI gene was expressed in maize.