Open AccessCloning and expression of the carbon monoxide dehydrogenase genes from Pseudomonas thermocarboxydovorans strain C2
Author(s) -
Black Gary W.,
Lyons Catherine M.,
Williams Edwin,
Colby John,
Kehoe Michael,
O'Reilly Catherine
Publication year - 1990
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1990.tb14005.x
Subject(s) - carbon monoxide dehydrogenase , cloning (programming) , gene , plasmid , microbiology and biotechnology , molecular cloning , blot , strain (injury) , protein subunit , southern blot , pseudomonas , genomic library , biology , expression vector , chemistry , recombinant dna , biochemistry , genetics , carbon monoxide , gene expression , bacteria , peptide sequence , anatomy , computer science , programming language , catalysis
SUMMARY Carbon monoxide dehydrogenase (CODH) from Pseudomonas thermocarboxydovorans strain C2 is composed of three non‐identical subunits. A gene library of C2 DNA in lambda vector L47.1 was generated and screened using anti‐CODH serum. Western blotting experiments revealed a protein which co‐migrated with and had the same immunological reaction as the large subunit of CODH in some of the clones isolated from the library. The coding region was pinpointed to a 4 kb fragment which was subcloned into plasmid. Western blotting experiments showed that all three subunits of CODH were coded for by the subclone. However, no CODH activity was detected.