【作者】 王丹；

【导师】 杨树林； 李大力；

【作者基本信息】 南京理工大学 ， 生物化工， 2002， 硕士

【摘要】 为获得转ubiC基因甘蓝，对植物基因工程载体的构建、甘蓝的组织培养以及基因转化条件进行了研究。通过PCR方法从大肠杆菌基因组中扩增得到了ubiC基因，扩增产物克隆到pUC118载体，转化大肠杆菌JM109。提取质粒，连接重组到经相同酶切的Ti质粒pBI121上，构建了植物表达载体pBI121-ubiC，并将其转入根癌农杆菌LBA4404中。同时，采用甘蓝无菌苗的下胚轴为外植体，以MS为基本培养基，分别用直接分化再生系统和愈伤再生系统诱导不定芽的分化，建立了高效稳定的再生系统，不定芽分化频率达80％以上。利用根癌农杆菌介导将ubiC基因转入甘蓝，通过预培养、侵染、共培养、脱菌培养、筛选转化体等步骤，最后获得抗性愈伤组织。更多还原

【Abstract】 The constuction of plant expression vector,tissue culture of cabbage and gene transformation conditions were studied to obtain ubiC transformants.ubiC gene was obtained from the Escherichia coli.by PCR amplication and was ligated to pUC118 vector. Subsequently, the recombined plasmid was transformed to the strain JM109 of Escherichia coli., which was picked up and was ligated to pBI121. As a result, the plant expression vector of pBI121-ubiC was constructed successfully. The hypocotyl segments of cabbage were used as the explants and cultured on a MS medium supplemented with plant hormone, which was established a cabbage regenerated system.The differentiation rate of adventitious buds was above 80%. In this study,ubiC gene was transformed to cabbage through Agrobacterium mediated system. And kanamycin-resistant callus tissue was obtained by pre-culture, infection co-culture and selection.更多还原