葡萄糖脱氢酶融合表达载体的构建

Construction of fusion expression vector of glucose dehydrogenase

目的为方便葡萄糖脱氢酶的纯化,构建葡萄糖脱氢酶融合表达载体。方法通过聚合酶链反应(PCR)从枯草杆菌基因组中扩增葡萄糖脱氢酶基因,与源序列比较分析后连接到融合表达载体pET22b,导入JM109诱导表达,测定葡萄糖脱氢酶酶液比活力。结果构建的葡萄糖脱氢酶融合表达载体中目的基因序列与枯草杆菌来源的该基因序列相同,且粗提酶液比活力高达375 U/mg。结论运用分子生物学技术获得葡萄糖脱氢酶基因,成功构建了葡萄糖脱氢酶的融合表达载体,为酶的后续纯化工作提供了条件。

Objective To facilitate the purification of glucose dehydrogenase and to construct fusion expression vector of glucose dehydrogenase.Methods Glucose dehydrogenase gene was amplified from the genome of Bacillus subtilis by PCR amplification,then compared sequence analysis with the source.Using the fusion expression vector pET22b with the glucose dehydrogenase gene,to introduce JM109 to induce expression and to determine the enzyme specific activity.Results The target gene from constructed glucose dehydrogenase gene fusion expression vector was the same as the source gene,and the rude enzyme specific activity was up to 375 U/mg.Conclusion The glucose dehydrogenase gene is gotten by using molecular biology techniques,and the fusion expression vector with the glucose dehydrogenase gene is successfully constructed,which provides the conditions for the purification of the enzyme.