Cry3a杀虫蛋白基因的合成优化及在JK-SH007中高效表达

发布时间：2018-04-30 19:27

本文选题：苏云金芽孢杆菌 + Crya基因　；参考：《中国生物工程杂志》2017年06期

【摘要】：人工合成优化Cry3a杀虫蛋白基因,并从大肠杆菌BL21(DE3)中克隆T7表达系统,通过同源重组克隆进PHKT2表达载体,将此表达载体导入吡咯伯克霍尔德氏菌JK-SH007,通过诱导表达,成功指导合成T7 RNA聚合酶且高效表达分子量为70k Da的Cry3a蛋白。通过粗提液对二龄榆蓝叶甲进行生物毒杀试验,结果表明粗提液具有高毒性,致死中浓度(LC_(50)=0.63(0.48-0.82)g/L)。成功构建的T7表达系统,可高效表达毒性的Cry3Aa蛋白,为进一步开发杀虫生防菌剂,建立外源基因表达技术平台奠定了基础。
[Abstract]:Cry3a insecticidal protein gene was synthesized and optimized. T7 expression system was cloned from Escherichia coli BL21 (DE3) and cloned into PHKT2 expression vector by homologous recombination. The expression vector was introduced into JK-SH007 by induction expression. T7 RNA polymerase was successfully synthesized and highly expressed Cry3a protein with a molecular weight of 70 kDa. The biological toxicity test of second instar Ulmus pumila was carried out by crude extract. The results showed that the crude extract was highly toxic, and the median lethal concentration was 0.63 ~ 0.63 ~ 0.48 ~ 0.82g / L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) ~ (-1) ~ (-1). The successfully constructed T7 expression system can express toxic Cry3Aa protein efficiently, which lays a foundation for the further development of insecticidal biocontrol agents and the establishment of foreign gene expression technology platform.
【作者单位】：南京林业大学林学院;湖州师范学院;
【基金】：国家社科基金(16@ZH005) 国家自然科学基金(31550004)资助项目
【分类号】：Q78;S476

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