CAJ | 학술논문

[目的]对缺失α1螺旋的Cry1Ac在大肠杆菌中的表达、纯化和活性进行分析。[方法]以苏云金芽孢杆菌HD-1中的质粒为模板,扩增Cry1Ac基因,并将其与pET28a载体连接,构建重组质粒。重组质粒转化并经IPTG诱导后,用SDS-PAGE检测目的蛋白的表达。提取目标蛋白,并涂在棉铃虫饲料表面,检测其杀虫活性。[结果]Cry1Acdelα1基因在大肠杆菌中能正常表达目标蛋白,但该目标蛋白是以包涵体形式存在。生测结果显示,在相同浓度下,活化的Cry1Ac对棉铃虫的致死率达75%以上,而Cry1Acdelα1只有10%左右。[结论]用大肠杆菌表达的Cry1Acdelα1对棉铃虫基本没有杀虫活性。
[목적]대결실α1라선적Cry1Ac재대장간균중적표체、순화화활성진행분석。[방법]이소운금아포간균HD-1중적질립위모판,확증Cry1Ac기인,병장기여pET28a재체련접,구건중조질립。중조질립전화병경IPTG유도후,용SDS-PAGE검측목적단백적표체。제취목표단백,병도재면령충사료표면,검측기살충활성。[결과]Cry1Acdelα1기인재대장간균중능정상표체목표단백,단해목표단백시이포함체형식존재。생측결과현시,재상동농도하,활화적Cry1Ac대면령충적치사솔체75%이상,이Cry1Acdelα1지유10%좌우。[결론]용대장간균표체적Cry1Acdelα1대면령충기본몰유살충활성。
[Objective] This study aimed to analyze the expression, purification and insecticidal activity of Cry1Ac without helix α-1 of domain I in E. coli. [Method] Plasmid of B. thuringiensis HD-1 was used as the template for amplification of Cry1Ac gene, which was linked with pET28a vector to construct the recombinant plasmid. After transformation and IPTG induction, expression of target protein was detected by using SDS-PAGE method. Target protein was extracted and coated on the surface of feed for H. armigera to determine the insecticidal activity. [Result] Cry1Acdela1 gene could normally express the target protein in E. coli. However, the target protein existed in the form of inclusion body. Results of bioassay analysis showed that under the same concentration, fatality rate of activated Cry1Ac toxin reached above 75%, while that of Cry1Acdelα1 was only about 10%. [Conclusion] E. coli-expressed Cry1Acdelα1 had no insecticidal activity against H. armigera.