[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.
Chi
Naiyu
N
College of Bioengineering, Dalian University, Dalian 116622, China. cny@dalianu.com
Liu
Changjiang
C
Liu
Yinghao
Y
Zhang
Qingfang
Q
Zheng
Xuefang
X
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D008970:Molecular Weight
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase