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通过花粉管通道法将pCMBIA3300-Cry1Iem载体中的Cry1Iem基因转入大豆品种“吉农28”中,对经过抗草铵膦筛选的转基因植株进行PCR检测,初步确定阳性植株,对转基因阳性植株进行Southern blotting检测,结果发现有5株出现杂交信号,证明Cry1Iem基因整合到受体的基因组中。利用实时荧光定量PCR测定T1代阳性植株的Cry1Iem基因在mRNA水平上的表达量,证明Cry1Iem基因在受体材料中获得了表达。
The Cry1Iem gene in pCMBIA3300-Cry1Iem vector was transferred into the soybean “Ji-nong28” by pollen tube channel method. The transgenic plants screened by glufosinate-resistant were detected by PCR and the positive plants were initially identified. blotting. As a result, five hybridization signals were found, indicating that the Cry1Iem gene was integrated into the genome of the receptor. The expression level of Cry1Iem gene in T1-positive plants was detected by real-time fluorescence quantitative PCR and the expression of Cry1Iem gene was confirmed in the receptor material.