Localization of the glnD gene on a revised map of the 200-kilobase region of the Escherichia coli chromosome

• Published in: Journal of bacteriology Vol. 174; no. 5; pp. 1702 - 1703
• Main Authors: VAN HEESWIJK, W, KUPPINGER, O, MERRICK, M, KAHN, D
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-03-1992
• Subjects: Bacteriology; Biological and medical sciences; Chromosome Mapping; Deoxyribonucleases, Type II Site-Specific - metabolism; DNA, Bacterial - metabolism; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genes, Bacterial - genetics; Genetics; Life Sciences; Microbiology; Other; Restriction Mapping; UDPglucose-Hexose-1-Phosphate Uridylyltransferase - genetics; Genetic mapping; Enzyme; Escherichia coli; Restriction map; Bacteria; Chromosome; Glutamine synthetase; Localization; Enterobacteriaceae
• DOI: 10.1128/jb.174.5.1702-1703.1992
• EISSN: 1098-5530
• ISSN: 0021-9193

To identify the physical location of glnD, we made use of a glnD::Tn10 insertion in strain RB9040 (3). Southern blotting of PstI-, EcoRI-, and EcoRI-SalI-digested genomic DNAs of RB9040 and its wild-type parent RB9010, using 21C8 phage DNA as a probe. This experiment located the Tn10 in strain RB9040 within a 2.8-kb PstI fragment, a 17-kb EcoRI fragment, and an 8-kb EcoRI-SalI fragment. The 8-kb EcoRI-SalI fragment was cloned from phage 21C8 into the Bluescript vector pBSK+, to obtain pWVH46. The physical map of pWVH46 was found to correspond with the physical map of an independently derived glnD-containing plasmid pXG204 (4a). pWVH46 was also found to complement the Gln super(-) bradytrophy of RB9040 (bradytrophy is defined as a leaky auxotrophy). Thus, the glnD gene was localized to the 2.8-kb segment separating dapD and map (4, 10; Fig. 2). Considering the molecular mass of uridylyl-transferase (95 kDa; 5), the glnD gene should be approximately 2.7 kb long.