Cloning and expression of a haloacid dehalogenase from Pseudomonas sp. strain 19 S.
A dehalogenase gene specifying the utilization of a variety of haloacids by Pseudomonas sp. Strain 19S has been cloned and expressed in E. coli. Our cloning strategy employed specific amplification of a fragment homologous to Pseudomonas dehalogenase gene by Polymerase Chain Reaction (PCR). The PCR amplicon successfully acted as a probe to detect the dehalogenase gene in the Southern Blot of the digested Pseudomonas total DNA. Corresponding fragments were cloned into pUC 18 vector and amplified in E. coli MV 1190. One clone with a substantial dehalogenation activity carried a recombinant plasmid containing a 5.5 kb insert.
Kocabiyik
S
S
Department of Biological Sciences, Middle East Technical University, Ankara, Turkey.
Aslan
B
B
Müller
R
R
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006867:Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics
D006867:Hydrolases; C034507:2-haloacid dehalogenase