Skip to main content
Log in

Cloning of ompF, the structural gene for an outer membrane pore protein of E. coli K12: Physical localization and homology with the phoE gene

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The ompF gene of Escherichia coli K12, which is the structural gene for an outer membrane pore protein has been cloned into the multicopy plasmid vector pACYC184. Expression of the cloned ompF gene results in overproduction of the OmpF protein and in strongly decreased production of the other two pore proteins, OmpC protein and PhoE protein, whereas production of OmpA protein is much less affected. Expression of the cloned ompF gene is still dependent on the ompB + gene product, but is hardly influenced by the osmolarity of the growth medium. The gene was localized on the hybrid plasmids by the analysis of in vitro constructed deletion plasmids and mutant plasmids generated by Tn5 insertions. Heteroduplex analysis revealed an extensive DNA homology of the ompF gene with the phoE gene. Since the direction of transcription of the phoE gene is known, the direction of transcription of the ompF gene could be deduced.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alphen W van, Lugtenberg B (1977) Influence of the osmolarity of the growth medium on the outer membrane protein pattern of Escherichia coli. J Bacteriol 131:623–630

    Google Scholar 

  • Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev 44:1–56

    Google Scholar 

  • Berg DE (1977) Insertion and excission of the transposable kanamycin resistance determinant Tn5. In: Bukhari AI, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids and episomes. Cold Spring Harbor Press, New York, pp 205–212

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1524

    Google Scholar 

  • Brown MGM, Weston A, Saunders JR, Humphreys GO (1979) Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth. FEMS Microbiol Lett 5:219–222

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Chen R, Krämer C, Schmidmayer W, Henning U (1979) Primary structure of major outer membrane protein I of Escherichia coli B/r. Proc Natl Acad Sci USA 76:5014–5017

    Google Scholar 

  • Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166

    Google Scholar 

  • Cosloy SD, Oishi M (1973) The nature of the transformation process in Escherichia coli K12. Mol Gen Genet 124:1–10

    Google Scholar 

  • Datta DB, Arden B, Henning U (1977) Major proteins of the Escherichia coli outer cell envelope membrane as bacteriophage receptors. J Bacteriol 131:821–829

    Google Scholar 

  • Davis RW, Simon M, Davidson N (1971) Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 21. Academic Press, New York, pp 413–428

    Google Scholar 

  • Davis RW, Davidson N (1968) Electron-microscope visualization of deletion mutations. Proc Natl Acad Sci USA 60:243–250

    Google Scholar 

  • Foulds J, Chai T (1978) New major outer membrane protein found in an Escherichia coli tolF mutant resistant to bacteriophage TuIb. J Bacteriol 133:1478–1483

    Google Scholar 

  • Hall MN, Silhavy TJ (1979) Transcriptional regulation of Escherichia coli K12 major outer membrane protein 1b. J Bacteriol 140:342–350

    Google Scholar 

  • Hasegawa Y, Yamada H, Mizushima S (1976) Interactions of outer membrane proteins 0–8 and 0–9 with peptidoglycan sacculus of Escherichia coli K12. J Biochem (Japan) 80:1401–1409

    Google Scholar 

  • Henning U, Schmidmayer W, Hindennach I (1977) Major proteins of the outer cell envelope membrane of Escherichia coli K12: multiple species of protein I. Mol Gen Genet 154:293–298

    Google Scholar 

  • Hondel CAMJJ van den, Keegstra W, Borrias WE, Arkel GA van (1979) Homology of plasmids in strains of unicellular cyanobacteria. Plasmid 2:323–333

    Google Scholar 

  • Ichihara S, Mizushima S (1978) Characterization of major outer membrane proteins 08 and 09 of Escherichia coli K12. Evidence that structural genes for the two proteins are different. J Biochem (Japan) 83:1095–1100

    Google Scholar 

  • Lugtenberg B, Boxtel R van, Verhoef C, Alphen W van (1978) Pore protein e of the outer membrane of Escherichia coli K12. FEBS Lett 96:99–105

    Google Scholar 

  • Lugtenberg B, Meijers J, Peters R, Hoek P van der, Alphen L van (1975) Electrophoretic resolution of the “major outer membrane protein” of Escherichia coli K12 into four bands. FEBS Lett 58:254–258

    Google Scholar 

  • Lugtenberg B, Peters R, Bernheimer H, Berendsen W (1976) Influence of cultural conditions and mutations on the composition of the outer membrane proteins of Escherichia coli. Mol Gen Genet 147:251–262

    Google Scholar 

  • Mutoh N, Nagasawa T, Mizushima S (1981) Specialized transducing bacteriophage lambda carrying the structural gene for a major outer membrane matrix protein of Escherichia coli K-12. J Bacteriol 145:1085–1090

    Google Scholar 

  • Nikaido H (1979) Nonspecific transport through the outer membrane. In: Inouye M (ed) Bacterial outer membranes: biogenesis and functions. Wiley-Interscience, New York, pp 361–407

    Google Scholar 

  • Ohsawa H, Maruo B (1976) Restoration by ribosomal protein S1 of the defective translation in a temperature-sensitive mutant of Escherichia coli K-12: characterization and genetic study. J Bacteriol 127:1157–1166

    Google Scholar 

  • Overbeeke N, Lugtenberg B (1980a) Expression of outer membrane protein e of Escherichia coli by phosphate limitation. FEBS Lett 112:229–232

    Google Scholar 

  • Overbeeke N, Lugtenberg B (1980b) Major outer membrane proteins of Escherichia coli strains of human origin. J Gen Microbiol 121:373–380

    Google Scholar 

  • Overbeeke N, Scharrenburg G van, Lugtenberg B (1980) Antigenic relationships between pore proteins of Escherichia coli K12. Eur J Biochem 110:247–254

    Google Scholar 

  • Rothstein SJ, Reznikoff WS (1981) The functional differences in the inverted repeats of Tn5 are caused by a single base pair non homology. Cell 23:191–199

    Google Scholar 

  • Sato T, Yura T (1979) Chromosomal location and expression of the structural gene for major outer membrane protein Ia of Escherichia coli K12 and of a homologous gene of Salmonella typhimurium. J Bacteriol 139:468–477

    Google Scholar 

  • Tanaku T, Weisblum B (1975) Construction of a colicin E1-R factor composite plasmid in vivo. J Bacteriol 121:354–362

    Google Scholar 

  • Tommassen J, Ley P van der, Lugtenberg B (1981) Genetic and biochemical characterization of an Escherichia coli K-12 mutant with an altered outer membrane protein. Antonic van Leeuwenhoek 47:325–337

    Google Scholar 

  • Tommassen J, Lugtenberg B (1980) Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase. J Bacteriol 143:151–157

    Google Scholar 

  • Tommassen J, Lugtenberg B (1981) Localization of phoE, the structural gene for outer membrane protein e in Escherichia coli K-12. J Bacteriol 147:118–123

    Google Scholar 

  • Tommassen J, Overduin P, Lugtenberg B, Bergmans H (1982) Cloning of phoE, the structural gene for the Escherichia coli K-12 phosphate limitation inducible outer membrane pore protein. J Bacteriol 149: (in press)

  • Verhoef C, Lugtenberg B, Boxtel R van, Graaff P de, Verhey H (1979) Genetics and biochemistry of the peptidoglycan-associated protein b and c of Escherichia coli K12. Mol Gen Genet 169:137–146

    Google Scholar 

  • Willetts NS, Clark AJ, Low KB (1969) Genetic location of certain mutations conferring recombination deficiency in Escherichia coli. J Bacteriol 97:244–249

    Google Scholar 

  • Yamamoto M, Nomura M, Ohsawa H, Maruo B (1977) Identification of a temperature-sensitive asparaginyl-transfer ribonucleic acid synthetase mutant of Escherichia coli. J Bacteriol 132:127–131

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by W. Arber

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tommassen, J., van der Ley, P., van der Ende, A. et al. Cloning of ompF, the structural gene for an outer membrane pore protein of E. coli K12: Physical localization and homology with the phoE gene. Molec Gen Genet 185, 105–110 (1982). https://doi.org/10.1007/BF00333798

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00333798

Keywords

Navigation