Summary
We have cloned the complete functional ompB locus of Salmonella typhimurium LT-2 into Escherichia coli K-12 using a cosmid vector and in vitro packaging into λ. The ompB locus of Salmonella was found to complement both envZ and ompR mutations in E. coli as well as an ompR mutation of Salmonella. The ompR part of the ompB locus was further subcloned into the multicopy plasmid pKN410 as a 1.3 kb fragment. This fragment coded for a single 28.5 kd protein corresponding to about 820 bp in length. Furthermore, the OmpR proteins of S. typhimurium and E. coli were shown to be structurally and functionally highly similar.
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Abbreviations
- SDS:
-
sedium dodecyl sulfate
- kb:
-
kilobase pairs
- bp:
-
base pairs
- kd:
-
kilodaltons
References
Amundsen SK, Neville ME (1979) Comparison of three procedures for isolating DNA from bacteria. Microbios 24:29–39
Bassford PJ, Diedrich DL, Schnaitman CA, Reeves P (1977) Outer membrane proteins of Escherichia coli. VI. Protein alteration in bacteriophage resistant mutants. J Bacteriol 131:608–622
Bochner BR, Huang H-C, Schieven GL, Ames BN (1980) Positive selection for loss of tetracycline resistance. J Bacteriol 143:926–933
Casadaban MJ (1976) Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage λ and Mu. J Mol Biol 104:541–555
Cleveland DW, Fisher SG, Kirschner MW, Laemmli UK (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 252:1102–1106
Clewell DB (1972) Nature of ColE1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676
Collins J, Brüning HJ (1978) Plasmids useable as gene-cloning vectors in an in vitro packaging by coliphage λ: “cosmids”. Gene 4:85–107
Datta DB, Kramer C, Henning U (1976) Diploidy for a structural gene specifying a major protein of the outer cell envelope membrane from E. coli K-12. J Bacteriol 128:834–841
Davies JK, Reeves P (1975) Genetics of resistance to colicin in Escherichia coli K-12: cross-resistance among colicins of group A. J Bacteriol 123:102–117
Fairbanks G, Steck TL, Wallach DFH (1971) Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617
Foulds J (1972) Purification and particle characterization of a bacteriocin in Serratia marcescen. J Bacteriol 110:1001–1009
Hall MN, Silhavy TJ (1979) Transcriptional regulation of Escherichia coli K-12 major outer membrane protein Ib. J Bacteriol 140:342–350
Hall MN, Silhavy TJ (1981a) The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli. J Mol Biol 146:23–43
Hall MN, Silhavy TJ (1981b) Genetic analysis of the ompB locus of Escherichia coli K-12. J Mol Biol 151:1–15
Hall MN, Silhavy TJ (1981c) Genetic analysis of the major outer membrane proteins of Escherichia coli. Annu Rev Genet 15:91–142
Hohn B, Collins J (1980) A small cosmid for efficient cloning of large DNA fragments. Gene 11:291–298
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Maloy SR, Nunn WD (1981) Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol 145:1110–1112
Laemmli UK (1970) Clearage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) New Biol 227:680–685
Lee DR, Schnaitman CA (1980) Comparison of outer membrane porin proteins produced by Escherichia coli and Salmonella typhimurium. J Bacteriol 142:1019–1022
Liljeström P, Määttänen PL, Palva ET (1982) Cloning of the regulatory locus ompB of Salmonella typhimurium LT-2. II. Identification of the envZ gene product, a protein involved in the expression of the porin proteins. Mol Gen Genet 188:190–194
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Nurminen M, Lounatmaa K, Sarvas M, Mäkelä PH, Nakae T (1976) Bacteriophage-resistant mutants of Salmonella typhimurium deficient in two outer membrane proteins. J Bacteriol 127:941–955
Osborn MJ, Wu HCP (1980) Proteins of the outer membrane of Gram negative bacteria. Annu Rev Microbiol 34:369–422
Palva ET (1978) Major outer membrane protein in Salmonella typhimurium induced by maltose. J Bacteriol 136:286–294
Palva ET (1979) Relationship between ompB genes of Escherichia coli and Salmonella typhimurium. FEMS Microbiol Lett 5:205–209
Palva ET, Liljeström P, Harayama S (1981) Cosmid cloning and transposon mutagenesis in Salmonella typhimurium using phage λ vehicles. Mol Gen Genet 181:153–157
Parkinson S (1976) cheA, cheB and cheC genes of Echerichia coli and their role in chemotaxis. J Bacteriol 126:758–770
Pugsley AP, Schnaitman CA (1978) Identification of 3 genes controlling production of new outer membrane pore proteins in Escherichia coli K-12. J Bacteriol 135:1118–1129
Randall LL, Hardy SJS, Josefsson L-G (1978) Precursors of three exported proteins in Escherichia coli. Proc Natl Acad Sci USA 75:1209–1212
Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693
Sarma V, Reeves P (1977) Genetic locus (ompB) affecting a major outer-membrane protein in Escherichia coli K-12. J Bacteriol 132:23–27
Sato T, Yura T (1979) Chromosomal location and expression of the structural gene for major outer membrane protein Ia of Escherichia coli K-12 and of the homologous gene of Salmonella typhimurium. J Bacteriol 139:468–477
Sutcliffe JG (1979) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90
Taylor RK, Hall MN, Enquist L, Silhavy TJ (1981) Identification of OmpR: a positive regulatory protein controlling expression of the major outer membrane porin proteins of E. coli K-12. J Bacteriol 147:255–258
Uhlin BE, Molin S, Gustafsson P, Nordström K (1979) Plasmid with temperature-dependent copy number for amplification of cloned genes and their products. Gene 6:91–106
Wandersman C, Moreno F, Schwartz M (1980) Pleiotropic mutations rendering Escherichia coli K-12 resistant to bacteriophage TP1. J Bacteriol 143:1374–1383
Wurtzel ET, Movva NR, Ross FL, Inouye M (1981) Two-step cloning of the Escherichia coli regulatory gene ompB, employing phage Mu. J Mol Appl Genet 1:61–69
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Liljeström, P., Määttänen, P.L. & Tapio Palva, E. Cloning of the regulatory locus ompB of Salmonella typhimurium LT-2. Mol Gen Genet 188, 184–189 (1982). https://doi.org/10.1007/BF00332673
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DOI: https://doi.org/10.1007/BF00332673