GENE FCBB CODING 4-CHLOROBENZOATE-COENZYME A DEHALOGENASE OF PSEUDOMONAS SP. DJ-12 AND RECOMBINANT PLASMID VECTOR THEREOF

摘要

PURPOSE: Provided is gene fcbB cloned from Pseudomonas sp. J-12. And a recombinant plasmid vector including the gene is also provided which can be inserted into bacteria to transform the strain to produce 4-chlorobenzoate-coenzyme A dehalogenase in large quantities which biologically dissolve aromatic compounds. CONSTITUTION: Gene fcbB is obtained by the next steps of: i) obtaining DNA from Pseudomonas sp. DJ-12 and cutting DNA with restriction enzyme Sau3A I; ii) linking DNA segments with cosmid pWE15 as a vector which is cut by BamH I at 16 deg.C for 16 hours; iii) transforming E. coli LE392 with the cosmid pWE15 having the segments; iv) selecting bacteria showing enzyme activity of dechlorination; and v) making many subclones from the bacteria. Cloned pKC1 is obtained from 40 kb Sau3A having gene fcbB and the gene fcbB in pKC1 plasmid is subcloned into pKC15 (36kb), pKC157 (22KB), and pKC158.