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通过根癌农杆菌介导,将抗虫基因-豇豆胰蛋白酶抑制剂(CowpeaTrypsinInhibitor,简称CpTI)基因导入花椰菜无菌苗的下胚轴和子叶中。卡那霉素(Kanamycin,简称Kan)的筛选浓度为15mg/L,抑制农杆菌生长的抗生素选用羧苄青霉素(Carbencillin,简称Carb),浓度为500mg/L。对所获得的14株Kan抗性植株进行PCR扩增,结果显示有8株为阳性;对PCR阳性植株进行Southern分子杂交检测,结果证明CpTI基因已被整合到花椰菜植株的基因组中。
The gene of Cowpea Trypsin Inhibitor (CpTI) was introduced into the hypocotyls and cotyledons of the cauliflower-free bacterium via Agrobacterium tumefaciens-mediated transformation. The screening concentration of Kanamycin (Kan) was 15 mg / L, and the antibiotic that inhibited the growth of Agrobacterium was selected to be Carbencillin (Carbacillin) at a concentration of 500 mg / L. The results of PCR amplification of the 14 Kan-resistant plants showed that 8 of them were positive. Southern blot analysis of PCR-positive plants showed that the CpTI gene had been integrated into the genome of cauliflower plants.