Abstract
The cloning of the gene for Escherichia coli PL-2 2-keto-3-deoxy-D-manno-octonate 8-phosphate synthetase is reported. Positive transformants showed an increase of approximately three-fold in specific activity of the enzyme both in E. coli and in Salmonella typhimurium as host cells. A subclone containing a 1.5-kilobase PvuII fragment overproduced active enzyme. Minicell experiments that allow the detection of plasmid encoded proteins revealed an insert-coded single protein band of 34 kilodaltons.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen-Hagge T., Lehmann V., Lüderitz O. Free flow electrophoresis as a tool for enrichment of mutants with temperature-dependent lethal mutations in lipid A synthesis. Eur J Biochem. 1985 Apr 1;148(1):21–27. doi: 10.1111/j.1432-1033.1985.tb08801.x. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lehmann V., Rupprecht E., Osborn M. J. Isolation of mutants conditionally blocked in the biosynthesis of the 3-deoxy-D-manno-octulosonic-acid--lipid-A part of lipopolysaccharides derived from Salmonella typhimurium. Eur J Biochem. 1977 Jun 1;76(1):41–49. doi: 10.1111/j.1432-1033.1977.tb11568.x. [DOI] [PubMed] [Google Scholar]
- Levy S. B. R factor proteins synthesized in Escherichia coli minicells: incorporation studies with different R factors and detection of deoxyribonucleic acid-binding proteins. J Bacteriol. 1974 Dec;120(3):1451–1463. doi: 10.1128/jb.120.3.1451-1463.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Munson R. S., Jr, Rasmussen N. S., Osborn M. J. Biosynthesis of lipid A. Enzymatic incorporation of 3-deoxy-D-mannooctulosonate into a precursor of lipid A in Salmonella typhimurium. J Biol Chem. 1978 Mar 10;253(5):1503–1511. [PubMed] [Google Scholar]
- Ray P. H. Purification and characterization of 3-deoxy-D-manno-octulosonate 8-phosphate synthetase from Escherichia coli. J Bacteriol. 1980 Feb;141(2):635–644. doi: 10.1128/jb.141.2.635-644.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rick P. D., Fung L. W., Ho C., Osborn M. J. Lipid A mutants of Salmonella typhimurium. Purification and characterization of a lipid A precursor produced by a mutant in 3-deoxy-D-mannooctulosonate-8-phosphate synthetase. J Biol Chem. 1977 Jul 25;252(14):4904–4912. [PubMed] [Google Scholar]
- Rick P. D., Osborn M. J. Isolation of a mutant of Salmonella typhimurium dependent on D-arabinose-5-phosphate for growth and synthesis of 3-deoxy-D-mannoctulosonate (ketodeoxyoctonate). Proc Natl Acad Sci U S A. 1972 Dec;69(12):3756–3760. doi: 10.1073/pnas.69.12.3756. [DOI] [PMC free article] [PubMed] [Google Scholar]