[Cloning and expression of the genes encoding glycerol dehydratase reactivase and identification of its biological activity].
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).
Li
Wen-Jun
WJ
Key Laboratory of Industrial Biotechnology Fujian Province, Hua Qiao University, Quanzhou 362021, China.
Fang
Bai-Shan
BS
Hong
Yan
Y
Wang
Xiao-Xia
XX
Lin
Jin-Xia
JX
Liu
Gui-Lan
GL
D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006836:Hydro-Lyases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction
D001426:Bacterial Proteins; D006836:Hydro-Lyases; C120061:glycerol dehydratase reactivating factor, Klebsiella pneumoniae