苏云金芽孢杆菌aiiA基因在毕赤酵母中的分泌表达  

Expression of Bacillus thuringiensis aiiA in Pichia pastoris

作　　者：毛惠民[1] 林丽玉[1] 杨彩云[1] 温真[1] 曾世涌[1] 杨梅[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》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)

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.

关 键 词：GS115 AIIA基因 PPIC9K 分泌表达 

分 类 号：S432.4[农业科学—植物病理学]