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草地学报  2015 

东方山羊豆GoGID基因表达载体构建及紫花苜蓿转化

DOI: 10.11733/j.issn.1007-0435.2015.01.026, PP. 167-172

Keywords: 东方山羊豆,赤霉素受体,紫花苜蓿,遗传转化

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Abstract:

根据已经克隆得到的东方山羊豆赤霉素受体(GoGID)基因,扩增编码区cDNA.以pBI121为基础载体,采用酶切连接法,构建植物超表达载体pBI121-GoGID.酶切鉴定表明目的基因已经正确插入载体中,超表达载体构建成功.采用CaCl2冻融法将重组载体转入农杆菌菌株中.以叶片为外植体,采用农杆菌介导的愈伤组织培养法,转化紫花苜蓿(Medicagesativa),得到抗性苗.对载体携带的nptⅡ基因、GUS基因进行PCR检测均成阳性,表明目的基因已成功导入紫花苜蓿基因组中.同时对转基因植株进行Southern-blot及RT-PCR检测,并均得到目的条带.本研究为进一步分析GoGID基因对紫花苜蓿生物量影响奠定了基础.

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