CAJ | 학술논문

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。
근거식물밀마자적편호성급사용빈솔，대소운금아포간균Cry1Ab야생형기인적편마구서렬진행우화화개조，개조후적Cry1Abm기인서렬여원시서렬동원성위66.2%，G+C함량유37.3%제고도62.7%。인공합성Cry1Abm기인，병장인공합성적개조후적Cry1Abm기인구건도원핵표체재체pET28b중，구건원핵표체재체pETAbm。장원핵표체재체전입대장간균BL21(DE3)중진행유도표체，용유도표체적단백진행사충(옥미명)실험。결과표명，해단백대유충구유흔강독성，유충적사망솔고체86.63%，동시，존활유충적생장발육야수도명현억제。해기인가이작위살충공정급배육전기인항충작물적후선기인。
The coding regions of wild Bacillus thuringiensis Cry1Ab was optimized and modified according to plant preferred codons and frequency. The modified Cry1Abm gene had 66.2 percent homology with the native cry1Ab gene, and G+C content was raised from 37.3% to 62.7%. The modified Cry1Abm gene was artificially synthesized. Modified Cry1Abm gene was cloned into prokaryotic expression vector pET28bm, to construct plasmid pETAbm. The protein expression in E. coli BL21(DE3) was confirmed by SDS-PAGE analysis. The results of insect assay with Asian corn borer showed the protein crude expression products of Cry1Abm gene in E. coli had a toxicity to corn borer. The mortality of insect larvae was accounted for 86.63%, and the growth of the survival larvae was seriously inhibited.