Cloning and characterization of hydrogenase genes from Azotobacter chroococcum |
| |
| Authors: | Karl H. Tibelius Robert L. Robson M. G. Yates |
| |
| Affiliation: | (1) AFRC Unit of Nitrogen Fixation, University of Sussex, BN1 9RQ Brighton, UK;(2) Present address: Department of Microbiology, Macdonald College, McGill University, Ste. Anne de Bellevue, H9X 1C0 Quebec, Canada |
| |
| Abstract: | Summary Genomic DNA from Azotobacter chroococcum was shown by DNA hybridization to contain sequences homologous to Rhizobium japonicum H2-uptake (hup) hydrogenase genes carried on the plasmid pHU1. Two recombinant cosmid clones, pACD101 and pACD102, were isolated from a gene library of A. chroococcum by colony hybridization and physically mapped. Each contained approximately 42 kb of insert DNA with approximately 27 kb of overlapping DNA. Further hybridization studies using three fragments from pHU1 (6 kb HindIII, 6.4 kb BglII and 5 kb EcoRI) showed that the hup-specific regions of R. japonicum and A. chroococcum are probably highly conserved. Weak homology to the hydrogenase structural genes from Desulfovibrio vulgaris (Hildenborough) was also observed. A 24 kb BamHI fragment from pACD102 subcloned into a broad host-range vector restored hydrogenase activity to several Hup- mutants of A. chroococcum. |
| |
| Keywords: | Hydrogenase genes Azotobacter chroococcum |
| 本文献已被 SpringerLink 等数据库收录！ |
|
