论文部分内容阅读
根据植物密码子的偏好性及使用频率,对苏云金芽孢杆菌Cry1Ab野生型基因的编码区序列进行优化和改造,改造后的Cry1Abm基因序列与原始序列同源性为66.2%,G+C含量由37.3%提高到62.7%。人工合成Cry1Abm基因,并将人工合成的改造后的Cry1Abm基因构建到原核表达载体pET28b中,构建原核表达载体pETAbm。将原核表达载体转入大肠杆菌BL21(DE3)中进行诱导表达,用诱导表达的蛋白进行饲虫(玉米螟)实验。结果表明,该蛋白对幼虫具有很强毒性,幼虫的死亡率高达86.63%,同时,存活幼虫的生长发育也受到明显抑制。该基因可以作为杀虫工程及培育转基因抗虫作物的候选基因。
According to the codon preference and frequency of plant, the coding region of Cry1Ab wild-type gene of B. thuringiensis was optimized and modified. The sequence of the modified Cry1Abm gene was 66.2% homologous to the original sequence and the G + C content was increased from 37.3 % To 62.7%. The Cry1Abm gene was artificially synthesized and the engineered Cry1Abm gene was constructed into the prokaryotic expression vector pET28b to construct the prokaryotic expression vector pETAbm. The prokaryotic expression vector was transformed into E. coli BL21 (DE3) for inducing expression, and the induced expression of the protein was used for feeding insects (Ostrinia furnacalis) experiment. The results showed that the protein was highly toxic to larvae, the larvae mortality rate as high as 86.63%, while the survival of larvae growth and development were also significantly inhibited. This gene can serve as a candidate gene for insecticidal engineering and breeding of transgenic insect-resistant crops.