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利用DNA合成仪人工合成了豇豆胰蛋白酶抑制剂(CpTI)的cDNA编码全序列。合成的基因经过克隆和序列分析后,克隆到植物高效表达载体上,并转化农杆菌,通过共转化的方法,将CpTI基因和经人工改造的苏云金芽孢杆菌(B.t)δ-内毒素基因共转化烟草,得到经PCR扩增并Southern-blotting验证的分别含有CpTI和B.t基因的植株以及同时含有CpTI和B.t基因的植株。利用棉铃虫幼虫进行的杀虫测试表明,转基因烟草和对照烟草相比具有明显的杀虫活性,同时转双基因的烟草和转单一基因的烟草相比具有增强的杀虫活性。
The full-length cDNA encoding cowpea trypsin inhibitor (CpTI) was synthesized by DNA synthesizer. After cloning and sequence analysis, the synthesized gene was cloned into the plant expression vector and transformed into Agrobacterium. The CpTI gene and the engineered Bacillus thuringiensis (Bt) δ-endotoxin gene were co-transformed by co-transformation Nicotiana tabacum, plants with CpTI and Bt genes respectively verified by PCR amplification and Southern-blotting, and plants containing both CpTI and Bt genes were obtained. Insecticidal tests using cotton bollworm larvae showed that transgenic tobaccos had significant insecticidal activity compared to control tobaccos while transgenic tobacco had enhanced insecticidal activity compared with tobacco-transgenics.