摘要
苏云金芽孢杆菌表达的Cry1Ia蛋白对鳞翅目和鞘翅目昆虫具有较好的杀虫活性。为便于后期评价不同的Cry1Ia蛋白,建立了Cry1Ia蛋白的表达与纯化体系。将一个cry1Ia基因的编码基因接入pET28aDel表达载体,并将其转化大肠杆菌BL21(DE3)star菌株。在大肠杆菌中诱导Cry1Ia蛋白的表达,并与Bt中的表达产物进行比较。最后,利用Cry1Ia蛋白C端包含的组氨酸标签,对大肠杆菌表达的Cry1Ia蛋白进行纯化。结果显示,大肠杆菌中Cry1Ia蛋白随着表达时间的延长,产物积累显著,并且其表达量高于Bt菌株,因此,其更适宜生产完整的Cry1Ia蛋白。对大肠杆菌表达的Cry1Ia进行纯化,获得了理想的结果。研究构建了Cry1Ia蛋白的大肠杆菌表达体系,并成功对其进行纯化,可为进一步研究此类蛋白的杀虫活性和机制奠定基础。
Cry1Ia proteins of Bacillus thuringiensis showed ideal insecticidal activity against Lepidoptera and Coleoptera pests. To evaluate different Cry1Ia proteins, the expression and purification system of Cry1Ia protein was established. A cry1Ia gene was inserted into the pET28aDel expression vector and transformed into Escherichia coli BL21(DE3) star strain. The expression of Cry1Ia protein was induced and were compared with that of Bt strain. Finally, the Esoherichia coli expressed Cry1Ia protein was purified by the histidine tag located at the C-terminus. The results showed that the Cry1Ia protein in Escherichia coli accumulated significantly with the prolongation of expression time, and the expression level was higher than that of Bt strain, so it was more suitable to produce the intact Cry1Ia protein. The constructed Cry1Ia protein could be purified successfully. In this study, the Escherichia coli expression and purification system of Cry1Ia protein was established which laid a foundation for further study of the insecticidal activity and mechanism of these proteins.
作者
郭小琴
钱红梅
郭星
高佳丽
张义茹
高建华
王兴春
GUO Xiaoqin;QIAN Hongmei;GUO Xing;GAO Jiali;ZHANG Yiru;GAO Jianhua;WANG Xingchun(College of Life Sciences,Shanxi Agricultural University,Taigu 030801,China;Institute of Agricultural Bioengineering,Shanxi Agricultural University,Taigu 030801,China)
出处
《山西农业科学》
2019年第5期748-750,769,共4页
Journal of Shanxi Agricultural Sciences
基金
国家自然科学基金项目(31601690)
山西省研究生教育创新项目(2018BY066)
关键词
苏云金芽孢杆菌
大肠杆菌
cry1Ia
蛋白表达
蛋白纯化
Bacillus thuringiensis
Escherichia coli
cry1Ia gene
protein expression
protein purification
