Molecular and General Genetics MGG

, Volume 181, Issue 3, pp 379–383 | Cite as

Molecular cloning of menaquinone biosynthetic genes of Escherichia coli K12

  • John R. Guest
  • Duncan J. Shaw


A tranducing phage carrying some of the genes (men) defining the early stages of menaquinone biosynthesis was isolated from a pool of recombinant lambda phages that had been constructed from R.HindIII digests of E. coli DNA and the corresponding insertion vector. The lesions of menB and menC mutants were complemented by the phage but menD mutants were transduced either at low frequencies or not at all. This indicates that the transducing phage contains functional menB and menC genes but that only part of the menD gene had been cloned. The phage (λG68) was accordingly disignated γmenCB(D). Studies with the transducing phage enabled earlier mapping data (Guest 1979) to be reinterpreted in favour of the gene order nalA.... menC..menB..menD.... purF. Restriction analyses established the presence of a bacterial DNA fragment (11.5 kb) linked by a R.HindIII target to the right arm of the γ genome but fused to the left arm of the vector. Hybridization studies confirmed that the cloned DNA was derived from a larger R.HindIII fragment (21 kb). A physical map of the men region was constructed and some flanking and overlapping fragments were identified.


Biosynthetic Gene Gene Order Hybridization Study Menaquinone Lambda Phage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Andrews S, Cox GB, Gibson F (1977) The anaerobic oxidation of dihydroorotate by Escherichia coli K-12. Biochem Biophys Acta 462:153–160Google Scholar
  2. Bachmann B, Low KB (1980) Linkage map of Escherichia coli K12, Edition 6. Microbiol Rev 1:1–56Google Scholar
  3. Borck K, Beggs JD, Brammar WJ, Hopkins, AS, Murray NE (1976) The construction in vitro of transducing derivatives of phage lambda. Mol Gen Genet 146:199–207Google Scholar
  4. Cole ST, Guest JR (1978) Studies with artificially-constructed fumarate reductase transducing phages (λfrdA). Soc Gen Microbiol Quarterly 6:43–44Google Scholar
  5. Cole ST, Guest JR (1980a) Genetic and physical characterization of lambda transducing phages (λfrdA) containing the fumarate reductase gene of Escherichia coli K12. Mol Gen Genet 178:409–418Google Scholar
  6. Cole ST, Guest JR (1980b) Amplification of fumarate reductase synthesis with λfrdA transducing phages and orientation of frdA gene expression. Mol Gen Genet 179:377–385Google Scholar
  7. Creaghan IT, Guest JR (1978) Succinate dehydrogenase-dependent nutritional requirement for succinate in mutants of Escherichia coli K12. J Gen Microbiol 107:1–13Google Scholar
  8. Daniels DL, DeWet JR, Blattner FR (1980) New map of bacteriophage lambda DNA. J Virol 33:390–400Google Scholar
  9. Guest JR (1977) Menaquinone biosynthesis: mutants of Escherichia coli K12 requiring 2-succinylbenzoate. J Bacteriol 130:1038–1046Google Scholar
  10. Guest JR (1979) Anaerobic growth of Escherichia coli K12 with fumarate as terminal electron acceptor. Genetic studies with menaquinone and fluoroacetate-resistant mutants. J Gen Microbiol 115:259–271Google Scholar
  11. Haddock BA, Jones CW (1977) Bacterial respiration. Bacteriol Rev 41:47–99Google Scholar
  12. Jabos, NJ, Jacobs JM (1978) Quinones as hydrogen carriers for a late step in anaerobic heme biosynthesis in Escherichia coli. Biochim Biophys Acta 544:540–546Google Scholar
  13. Krörger A (1977) Phosphorylative electron transport with fumarate and nitrate as terminal hydrogen acceptors. In: Haddock BA, Hamilton WA (eds) Microbial energetics, Symp Soc gen Microbiol, Vol 27. Cambridge University Press, Cambridge, p 61Google Scholar
  14. Kröger A (1978) Fumarate as terminal acceptor of phosphorylative electron transport. Biochim Biophys Acta 505:129–145Google Scholar
  15. Lambden PR, Guest JR (1976) Mutants of Escherichia coli K12 unable to use fumarate as an anerobic electron acceptor. J Gen Microbiol 97:145–160Google Scholar
  16. Meganathan R, Bentley R (1979) Menaquinone (vitamin K2) biosynthesis: conversion of o-succinylbenzoic acid to 1,4-dihydroxy-2-naphthoic acid by Mycobacterium phlei enzymes. J Bacteriol 140:92–98Google Scholar
  17. Meganathan R, Folger T, Bentley R (1980) Conversion of o-succinyl-benzoate to 1,4-dihydroxy-2-naphthoate by extracts of Micrococcus luteus. Biochemistry 19:785–789Google Scholar
  18. Murray NE, Manduca de Ritis P, Foster LA (1973) DNA targets for the E. coli K restriction system analysed genetically in recombinants between phages phi80 and lambda. Mol Gen Genet 120:261–280Google Scholar
  19. Murray NE, Brammar WJ, Murray K (1977) Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet 150:53–61Google Scholar
  20. Shaw DJ, Guest JR (1981) Molecular cloning of the fnr gene of Escherichia coli K12. Mol Gen Genet 181:95–100Google Scholar
  21. Shineberg B, Young IG (1976) Biosynthesis of bacterial menaquinones: the membrane-associated 1,4-dihydroxy-2-naphthoate octaprenyl-transferase of Escherichia coli. Biochemistry 15:2754–2758Google Scholar
  22. Wilson GG, Murray NE (1979) Molecular cloning of the DNA ligase gene from phage T4. I. Characterization of the recombinants. J Mol Biol 132:471–491Google Scholar
  23. Young IG (1975) Biosynthesis of bacterial quinones: menaquinone mutants of Escherichia coli. Biochemistry 14:399–406Google Scholar

Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • John R. Guest
    • 1
  • Duncan J. Shaw
    • 1
  1. 1.Department of MicrobiologyUniversity of SheffieldSheffieldUK

Personalised recommendations