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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 aiiA gene in GenBank, aiiA gene was amplified from Bacillus thuringiensis genome, then a recombinant plasmid pPIC9K-aiiA was constructed and transformed into Pichia pastoris GS115 after linearization to obtain the recombinant engineering bacterium GS115 / pPIC9K-aiiA 1% methanol was induced by SDS-PAGE and Western blotting.The results showed that the expressed protein had biological activity and good anti-tumor activity Disease ability.