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Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects

热3 duanxiaohong 添加于 2010-4-22 21:06 | 1971 次阅读 | 0 个评论

作者
Jingjing Liu , Guixin Yan , Changlong Shu , Can Zhao , Chunqin Liu , Fuping Song, Lin Zhou , Junlan Ma, Jie Zhang , Dafang Huang
摘要
A newly engineered strain, denominated BIOT185,was constructed through integrating the cry8Ca2 gene intothe endogenous plasmid of BT185 (contains cry8Ea1) byhomologous recombination. The thermosensitive plasmidvector was eliminated by the rising temperature of recombinant cultures. No antibiotic gene or other unnecessary geneswere introduced to the new strain. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blot analysis demonstrated that the cry8Ca2 gene was expressed normally and produced a 130-kDa protein in the BIOT185 strain.Bioassay results showed that the new strain had high toxicity to the pests Anomala corpulenta and Holotrichia parallela, which often damage the same fields.
详细资料
- 文献种类: Journal Article
学科领域生物医药 » 生物学
附件
2010.pdf

duanxiaohong 的文献笔记订阅

【原创】该文章构建了一种对鞘翅目害虫有高毒性的新型BT工程菌BIOT185, 构建的过程主要是：把一段4.3kb的cry8Ca2的核心编码区，后面连个cry3Aa7的启动子，两侧同时加上一个tnp167B 后，将其插入到pMAD载体的MCS区中，（MCS区先用MluI 和 NcoI切去），改造好的载体被命名PMCS，然后利用同源重组的方法将cry8Ca2基因导入到BT185的内源质粒中，巧妙的是cry8Ca2的载体是热敏感的，重组后可通过高温将其去除，从而使我们新构建的BT菌株中不含一些不必要的基因和抗生素等！通过跑SDS-PAGE和免疫印迹分子可看出我们的cry8Ca2基因能正常表达、产生130KDa的蛋白，然后作者又通过抗虫活性测定实验证明该蛋白确实对鞘翅目害虫有很强的杀虫毒性。很不错的文章哦，相信这个新的工程菌株以后一定会被广泛地应用于生物杀虫剂中！

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