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以大豆Williams 82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Uid A(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2 min时,报告基因GUS的瞬时表达效率达到最高;在0.3~0.6 mg·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。
Using the mature embryo of soybean Williams 82 as the experimental material, the ultrasonic-assisted transformation of Agrobacterium tumefaciens was established. The effect of different ultrasonic treatment time on the transient expression of reporter gene Uid A (GUS) and the effect of different glufosinate concentrations Effect of Soybean Transformation Efficiency. The results showed that the transient expression efficiency of reporter gene GUS reached the highest at 2 min after ultrasonic treatment. The number of positive transgenic plants was the highest at 0.3 ~ 0.6 mg · L-1 glufosinate gradient screening concentration. The transgenic lines were obtained by PCR detection and herbicide Basta smear resistance detection; real-time PCR detection showed that the target gene expression of transgenic plants reached 1.7 to 42 times the wild-type, Southern hybridization test showed that exogenous genes Has been integrated into the soybean genome.