编码1,3-丙二醇氧化还原酶基因的克隆和表达

采用PCR法克隆了巴氏梭菌（Clostridium pasteurianum）CpN86菌株编码1,3丙二醇氧化还原酶基因（dhaT基因）；完成了dhaT基因测序、表达载体构建和在大肠杆菌中表达；分离和纯化了dhaT基因表达的重组蛋白。实验结果：（1）PCR法克隆的dhaT基因和肺炎克雷伯氏菌Klebsiella pneumoniae菌株dhaT基因的序列同源性为829%；（2）dhaT基因表达蛋白的酶活为108U/mg；（3）dhaT基因表达的蛋白分子量为43kD；（4）Western blot确定了dhaT基因表达的蛋白和 CpN86菌株天然蛋白有相同的抗原反应。

Cloning and Expressing of 1,3-propanediol Oxidoreductase-encoding Gene

Based on what mentioned above, the gene encoding 1,3-propanediol Oxidoreductase (dhaT) in Clostridium pasteurianum CpN-86 was cloned by using PCR method. The sequence, expressing vector construction and its expression of dhaT in E. coli were accomplished respectively. The recombination protein expressed by dhaT was also isolated and purified. The experimental results showed that: (1) The homogeneity of dhaT cloned by PCR and that in klebsiella pneumoniae strain was 82.9%; (2)The enzymatic activity of the protein expressed by cloned dhaT was 108 microM/ mg; (3)The molecular weight of the protein was 43 kD; (4) The protein expressed by dhaT has the same antigenicity as the natural protein of CpN-86 through Western blotting.