Cloning and expression of the genes encoding glycerol dehydratase reactivase and identification of its biological activity.
- Author:
Wen-Jun LI
1
;
Bai-Shan FANG
;
Yan HONG
;
Xiao-Xia WANG
;
Jin-Xia LIN
;
Gui-Lan LIU
Author Information
1. Key Laboratory of Industrial Biotechnology Fujian Province, Hua Qiao University, Quanzhou 362021, China.
- Publication Type:Journal Article
- MeSH:
Bacterial Proteins;
genetics;
isolation & purification;
metabolism;
Cloning, Molecular;
Electrophoresis, Polyacrylamide Gel;
Escherichia coli;
genetics;
Gene Expression;
Genetic Vectors;
genetics;
Hydro-Lyases;
genetics;
isolation & purification;
metabolism;
Plasmids;
genetics;
Polymerase Chain Reaction
- From:
Chinese Journal of Biotechnology
2006;22(6):950-955
- CountryChina
- Language:Chinese
-
Abstract:
The gdrA, gdrB gene coding glycerol dehydratase reactivase factor were amplified by using the genomic DNA of Klebsiella pneumoniae as the template. The gdrA and gdrB were inserted in pMD-18T to yield the recombinant cloning vector pMD-gdrAB. After the DNA sequence was determined, the gdrAB gene was subcloned into expression vector pET-28a(+) to yield the recombinant expression vector pET-28gdrAB. Under screening pressure by ampicillin and kanamycin simultaneously, the activity of glycerol dehydratase reactivase was characterized by coexpression of pET-32gldABC, which carry the gldABC gene encoding glycerol dehydratase, and pET-28gdrAB in E. coli BL21(DE3).
