摘要
根据GenBank中aiiA基因设计引物,以苏云金芽孢杆菌基因组为模板扩增出aiiA基因后构建出pPIC9K-aiiA重组表达载体,线性化后转化毕赤酵母GS115,获得重组工程菌GS115/pPIC9K-aiiA,以体积分数为1%的甲醇进行诱导表达.表达产物经SDS-PAGE及Western blotting分析显示表达的蛋白具有较好的免疫特异性.抗病性实验显示表达的目的蛋白具有生物学活性和良好的抗病能力.
According to the aiiA gene in GenBank, a pair of primers was designed and used in a PCR to amplify the alia gene from the genome of Bacillus thuringiensis. The puri-fied PCR product was inserted into pPIC9K, the recombinant expressed vector pPIC9K-aiiA was constructed. Then the recombinant plasmid was transformed into Pichia pastoris GSl15 by electroporation after linearization. After that, the recombinant yeast GSllS/pPIC9K-aiiA was obtained, which was then induced to express the recombinant AiiA protein with methanol at the volume fraction of 1%. The results of SDS-PAGE analysis and Western blot-ting showed that the recombinant AiiA protein was expressed successfully in the yeast. Fur-thermore, the protein had both biological activity and disease resistant capability by resis-tance experiment.
出处
《福建师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2012年第5期113-118,共6页
Journal of Fujian Normal University：Natural Science Edition
基金
福建省自然科学基金资助项目(2012J01123)
