Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Feb;129(2):1078–1090. doi: 10.1128/jb.129.2.1078-1090.1977

Isolation and characterization of lambda pleu bacteriophages.

M G Davis, J M Calvo
PMCID: PMC235049  PMID: 320178

Abstract

In the Escherichia coli lysogen HfrH73 described by Shimada et al. (1973), none of the enzymes coded for by the leucine operon is synthesized due to an insertion of phage lambda into cistron leuA. The orientation of lambda in the chromosome is ara leuDCB lambda JAN leuA. After heat induction of the lysogen, plaque-forming transducing phages of two types are formed at low frequency. One type (e.g., lambda pleu9) transduces leuD, leuC, and leuB strains to prototrophy. The other type (e.g., lambda pleu 13) transduces leuA strains to prototrophy. lambda pleu 13 forms lysogens at low frequency (about 0.2%) by integration into the leucine operon. These lysogens are unstable, segregating phage-sensitive clones at high frequency (about 1%). Phages carrying different portions of the leucine operon were formed by aberrant excision after heat induction of strain CV437 (leuA371 lambda pleu13). A phage carrying the entire leucine operon (lambda K2) was constructed by a cross between lambda pleu9 and lambda pleu13. An analysis of leucine-forming enzyme levels in strains lysogenized with lambdaK2 indicated that leuO and leuP are present and functional in lambda K2. leu-specific messenger ribonucleic acid from E. coli hybridizes to the heavy (r) strand of lambdaK2. The leucine operon of lambda G4 pleuABCD (an S7 derivative of lambda K2) exists intact on a 7.3 x 10(6)-dalton fragment (lambdaG4EcoRI-B) generated by cleavage with endonuclease EcoRI. Heteroduplexes formed between lambda G4 and lambda show a 5.4 x 10(6)-dalton piece of bacterial deoxyribonucleic acid (DNA) replacing a 4.5 x 10(6)-dalton piece of lambda DNA starting at 0.46 fractional unit on the map of lambda. Fragment lambda G4EcoRI-B has about 0.6 x 10(6) daltons of lambda DNA from the b2 region at one end and about 1.4 x 10(6) daltons of lambda DNA from the int region at the other end.

Full text

PDF
1078

Images in this article

1087Image
on p.1087
1088Image
on p.1088

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burns R. O., Calvo J., Margolin P., Umbarger H. E. Expression of the leucine operon. J Bacteriol. 1966 Apr;91(4):1570–1576. doi: 10.1128/jb.91.4.1570-1576.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Calvo J. M., Bartholomew J. C., Stieglitz B. I. Fluorometric assay of enzymatic reactions involving acetyl Coenzyme A in aldol condensations. Anal Biochem. 1969 Apr 4;28(1):164–181. doi: 10.1016/0003-2697(69)90168-7. [DOI] [PubMed] [Google Scholar]
  3. Calvo J. M., Freundlich M., Umbarger H. E. Regulation of branched-chain amino acid biosynthesis in Salmonella typhimurium: isolation of regulatory mutants. J Bacteriol. 1969 Mar;97(3):1272–1282. doi: 10.1128/jb.97.3.1272-1282.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Germaine G. R., Rogers P. Role of gal repressor depletion in lambda-dg transduction escape synthesis. J Mol Biol. 1970 Jan 28;47(2):121–135. doi: 10.1016/0022-2836(70)90334-7. [DOI] [PubMed] [Google Scholar]
  5. Helling R. B., Goodman H. M., Boyer H. W. Analysis of endonuclease R-EcoRI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J Virol. 1974 Nov;14(5):1235–1244. doi: 10.1128/jvi.14.5.1235-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ikemura T., Dahlberg J. E. Small ribonucleic acids of Escherichia coli. I. Characterization by polyacrylamide gel electrophoresis and fingerprint analysis. J Biol Chem. 1973 Jul 25;248(14):5024–5032. [PubMed] [Google Scholar]
  7. Klingmüller W., Krell K., Shimada K. Genetic analysis of bacteriophage lambda strains which transduce the genes for leucine biosynthesis. Mol Gen Genet. 1973 Oct 16;126(1):1–6. doi: 10.1007/BF00333476. [DOI] [PubMed] [Google Scholar]
  8. Konrad B., Kirschbaum J., Austin S. Isolation and characterization of phi80 transducing bacteriophage for a ribonucleic acid polymerase gene. J Bacteriol. 1973 Nov;116(2):511–516. doi: 10.1128/jb.116.2.511-516.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lindahl G., Sironi G., Bialy H., Calendar R. Bacteriophage lambda; abortive infection of bacteria lysogenic for phage P2. Proc Natl Acad Sci U S A. 1970 Jul;66(3):587–594. doi: 10.1073/pnas.66.3.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Neubauer Z., Calef E. Immunity phase-shift in defective lysogens: non-mutational hereditary change of early regulation of lambda prophage. J Mol Biol. 1970 Jul 14;51(1):1–13. doi: 10.1016/0022-2836(70)90265-2. [DOI] [PubMed] [Google Scholar]
  12. Reader R. W., Siminovitch L. Lysis defective mutants of bacteriophage lambda: genetics and physiology of S cistron mutants. Virology. 1971 Mar;43(3):607–622. doi: 10.1016/0042-6822(71)90286-8. [DOI] [PubMed] [Google Scholar]
  13. Reznikoff W. S., Michels C. A., Cooper T. G., Silverstone A. E., Magasanik B. Inhibition of lacZ gene translation initiation in trp-lac fusion strains. J Bacteriol. 1974 Mar;117(3):1231–1239. doi: 10.1128/jb.117.3.1231-1239.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  15. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. doi: 10.1016/0022-2836(72)90443-3. [DOI] [PubMed] [Google Scholar]
  16. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. II. Mutations induced by bacteriophage lambda in Escherichia coli K12. J Mol Biol. 1973 Oct 25;80(2):297–314. doi: 10.1016/0022-2836(73)90174-5. [DOI] [PubMed] [Google Scholar]
  17. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. III. The components of the secondary attachment sites. J Mol Biol. 1975 Apr 25;93(4):415–429. doi: 10.1016/0022-2836(75)90237-5. [DOI] [PubMed] [Google Scholar]
  18. Sly W. S., Rabideau K. Mechanism of lambda-c17cI virulence. J Mol Biol. 1969 Jun 28;42(3):385–400. doi: 10.1016/0022-2836(69)90231-9. [DOI] [PubMed] [Google Scholar]
  19. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  20. Zubay G. In vitro synthesis of protein in microbial systems. Annu Rev Genet. 1973;7:267–287. doi: 10.1146/annurev.ge.07.120173.001411. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES