【摘 要】
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该论文研究了植酸酶基因在工程菌中的表达及初步改造.以含有植酸酶基因的PMD质粒为模板,利用PCR方法扩增出植酸酶基因片段,把表达载体pTrcHis2B和PCR产物进行重组,构建成功pT
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该论文研究了植酸酶基因在工程菌中的表达及初步改造.以含有植酸酶基因的PMD质粒为模板,利用PCR方法扩增出植酸酶基因片段,把表达载体pTrcHis2B和PCR产物进行重组,构建成功pTrcHis2B-phyA质粒,转入大肠杆菌JM109中进行发酵培养.对工程菌发酢产生的蛋白质进行了SDS-PAGE分析.通过error-pronePCR法对植酸酶基因进行体外随机突变,与载体pTrcHis2B重组后转入大肠杆菌中进行表达,筛选出热稳定性较高的菌株.测定突变株酶活与非突变株相近,热稳定性稍有提高,在65℃下30分钟后酶活剩余40﹪,提高30﹪.
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