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大豆科学  2014 

超声波辅助处理农杆菌介导大豆胚尖转化转基因植株的获得和分子鉴定

DOI: 10.11861/j.issn.1000-9841.2014.06.0808, PP. 808-814

Keywords: 大豆,胚尖,农杆菌转化,超声波辅助处理

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Abstract:

以大豆Williams82成熟胚尖为试验材料,建立了超声波辅助处理的大豆胚尖农杆菌转化体系,研究了不同超声波处理时间对报告基因UidA(GUS)瞬间表达的影响和不同草铵膦浓度对大豆转化效率的影响。结果表明:在超声波处理2min时,报告基因GUS的瞬时表达效率达到最高;在0.3~0.6mg.L-1草铵膦梯度筛选浓度下,阳性转基因植株的数量最多。对于所获得的转基因株系进行了PCR检测和除草剂Basta涂抹抗性检测;实时荧光定量PCR检测显示转基因植株的目的基因的表达量达到野生型的1.7~42倍,Southern杂交检测表明外源基因已经整合到大豆基因组中。

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