利用酵母磷酸甘露糖异构酶基因作为拟南芥遗传转化的筛选标记

Phosphomannose isomerase gene from Saccharomyces cerevisiae as a selectable marker for Arabidopsis gene transformation

利用PCR反应克隆来自酿酒酵母的磷酸甘露糖异构酶(PMI)基因pmi,通过农杆菌介导的花序侵染法将PMI的基因导入受体拟南芥中,将转化后的拟南芥种子放在质量浓度为1g/L的甘露糖MS培养基上进行筛选培养,共得到抗性拟南芥植株4株。PCR反应证明酵母PMI的基因pmi已经整合到拟南芥基因组中,RT-PCR检测证明该基因的内含子能够在拟南芥中被正确剪切,同时通过GUS化学组织染色方法证明了伴随转化的GUS基因gus在拟南芥中得以表达,以上研究结果说明酵母PMI/甘露糖系统可以作为拟南芥遗传转化的筛选标记。

Phosphomannose isomerase （PMI） gene from Saccharomyces cerevisiae was cloned by PCR,and transformed into Arabidopsis via floral dip method. The seeds were collected and germinated on selecting MS medium which mannose concentration was 1 g/L. Four positive plants were obtained. PCR for genome DNA and RT-PCR for RNA proved the PMI gene had integrated into the genome of Arabidopsis and was transcribed. The intron of yeast pmi was removed successfully in Arabidopsis. GUS activity also was detected in the transgenic plants. All the results means PMI gene from yeast can be used as a selectable marker in Arabidopsis transformation.