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草业学报  2014 

CryⅢA基因植物表达载体构建及马铃薯遗传转化

DOI: 10.11686/cyxb20140130, PP. 248-256

Keywords: In-Fusion,CryⅢ,A基因,表达载体,马铃薯,遗传转化

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Abstract:

利用In-Fusion技术将酶切位点不匹配、目的基因内部含载体酶切位点的CryⅢA基因和植物表达载体pBI121相连接,分别成功构建了组成型启动子CaMV35S和马铃薯茎叶特异表达启动子ST-LS1驱动的CryⅢA基因植物表达载体。利用冻融法将2个重组质粒转入根癌农杆菌LBA4404,转化马铃薯栽培品种陇薯3号和甘农薯2号获得了转化植株,经卡那霉素筛选和PCR检测,共有10株阳性植株CryⅢA基因成功整合到马铃薯基因组中。经实时荧光定量PCR检测表明,CryⅢA基因在CaMV35S驱动的CryⅢA转基因植株的根、茎、叶中均有表达,且在叶中表达量较高,茎和根次之;在马铃薯茎叶特异表达启动子ST-LS1驱动的CryⅢA转基因植株中,在根中未见表达,仅在茎、叶中表达,且在叶中表达量较高。

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