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大豆科学  2015 

GmAOC3-基因转化载体构建及转化大豆的初步研究

DOI: 10.11861/j.issn.1000-9841.2015.04.0588, PP. 588-596

Keywords: 大豆,丙二烯氧化物环化酶,载体构建,遗传转化,分子检测

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Abstract:

选用内切酶SacⅠ和MluⅠ切割目标基因和载体,构建了携带-bar-基因的重组载体pBA002-GmAOC3,选用2个大豆品种(Jack和南农88-1)和2种外植体(子叶节和整个子叶节),研究大豆品种和外植体对根癌农杆菌介导的子叶节转化-GmAOC3-基因的影响。结果表明:外植体为整个子叶节的大豆品种Jack的出芽率和转化率最高,分别为79.5%和2.27%。经QuickStixPAT/bar-基因试纸条检测、目标基因的分子检测和草丁膦抗性鉴定,共得到转-GmAOC3-基因的T0代阳性株系3株,T1代阳性转基因大豆6株,其中5株以Jack品种为受体的转基因大豆-GmAOC3-基因的表达量均显著高于非转基因植株,荧光定量拷贝数检测结果显示,4株转基因单株为单拷贝,2株为双拷贝。

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