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. 1971 Oct;8(4):551–563. doi: 10.1128/jvi.8.4.551-563.1971

Electron Microscopic Evidence for Linear Insertion of Bacteriophage MU-1 in Lysogenic Bacteria

J Martuscelli 1,2,1, A L Taylor 1,2, D J Cummings 1,2, V A Chapman 1,2, S S DeLong 1,2, L Cañedo 1,2,1
PMCID: PMC376228  PMID: 4943078

Abstract

Temperate bacteriophage Mu-1 was used to generate a lysogenic derivative of the F′lac episome of Escherichia coli. Intact, covalently circular molecules of F′lac and lysogenic F′lac Mu+ deoxyribonucleic acid (DNA) were isolated and examined by electron microscopy. The mean contour lengths of F′lac and F′lac Mu+ molecules were 37.6 ± 0.4 μm and 53.2 ± 0.4 μm, respectively. The mean difference, 15.6 μm, is similar to the mean contour length of 12.9 ± 0.1 μm obtained for linear DNA molecules released by osmotic shock from mature phage Mu-1 virions. These results provide direct physical evidence that phage Mu-1 integrates by linear insertion of its genome into the DNA of lysogenic host bacteria. Chemical and physical analyses of phage Mu-1 DNA indicate that it is similar to E. coli DNA in respect of gross base composition, buoyant density, and melting temperature.

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Selected References

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  1. ADELBERG E. A., BURNS S. N. Genetic variation in the sex factor of Escherichia coli. J Bacteriol. 1960 Mar;79:321–330. doi: 10.1128/jb.79.3.321-330.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ahmad S. I., Pritchard R. H. A map of four genes specifying enzymes involved in catabolism of nucleosides and deoxynucleosides in Escherichia coli. Mol Gen Genet. 1969 Aug 15;104(4):351–359. doi: 10.1007/BF00334234. [DOI] [PubMed] [Google Scholar]
  3. Ansz H. S., Zandberg J., van de Pol J. H., van Bruggen E. F. Circular DNA from Shigella paradysenteriae. Eur J Biochem. 1969 Jun;9(2):156–159. doi: 10.1111/j.1432-1033.1969.tb00589.x. [DOI] [PubMed] [Google Scholar]
  4. Bazaral M., Helinski D. R. Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J Mol Biol. 1968 Sep 14;36(2):185–194. doi: 10.1016/0022-2836(68)90374-4. [DOI] [PubMed] [Google Scholar]
  5. CARO L. G. THE MOLECULAR WEIGHT OF LAMBDA DNA. Virology. 1965 Feb;25:226–236. doi: 10.1016/0042-6822(65)90201-1. [DOI] [PubMed] [Google Scholar]
  6. CHARGAFF E., LIPSHITZ R., GREEN C., HODES M. E. The composition of the deoxyribonucleic acid of salmon sperm. J Biol Chem. 1951 Sep;192(1):223–230. [PubMed] [Google Scholar]
  7. Cummings D. J., Kusy A. R., Chapman V. A., DeLong S. S., Stone K. R. Characterization of T-even bacteriophage substructures. I. Tail fibers and tail tubes. J Virol. 1970 Oct;6(4):534–544. doi: 10.1128/jvi.6.4.534-544.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cummings D. J., Mondale L. Density-gradient banding of denatured deoxyribonucleic acid in cesium sulphate. Biochim Biophys Acta. 1966 Jul 13;120(3):448–453. doi: 10.1016/0926-6585(66)90311-6. [DOI] [PubMed] [Google Scholar]
  9. Freifelder D., Meselson M. Topological relationship of prophage lambda to the bacterial chromosome in lysogenic cells. Proc Natl Acad Sci U S A. 1970 Jan;65(1):200–205. doi: 10.1073/pnas.65.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Freifelder D. Studies with Escherichia coli sex factors. Cold Spring Harb Symp Quant Biol. 1968;33:425–434. doi: 10.1101/sqb.1968.033.01.049. [DOI] [PubMed] [Google Scholar]
  11. Hane M. W., Wood T. H. Escherichia coli K-12 mutants resistant to nalidixic acid: genetic mapping and dominance studies. J Bacteriol. 1969 Jul;99(1):238–241. doi: 10.1128/jb.99.1.238-241.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hirota Y. THE EFFECT OF ACRIDINE DYES ON MATING TYPE FACTORS IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1960 Jan;46(1):57–64. doi: 10.1073/pnas.46.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jordan E., Saedler H., Starlinger P. O0 and strong-polar mutations in the gal operon are insertions. Mol Gen Genet. 1968;102(4):353–363. doi: 10.1007/BF00433726. [DOI] [PubMed] [Google Scholar]
  14. KELLENBERGER G., ZICHICHI M. L., WEIGLE J. J. Exchange of DNA in the recombination of bacteriophage lambda. Proc Natl Acad Sci U S A. 1961 Jun 15;47:869–878. doi: 10.1073/pnas.47.6.869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. KLEINSCHMIDT A. K., LANG D., JACHERTS D., ZAHN R. K. [Preparation and length measurements of the total desoxyribonucleic acid content of T2 bacteriophages]. Biochim Biophys Acta. 1962 Dec 31;61:857–864. [PubMed] [Google Scholar]
  16. Kemp C. L., Howatson A. F., Siminovitch L. Electron microscopy studies of mutants of lambada bacteriophage. I. General description and quantitation of viral products. Virology. 1968 Nov;36(3):490–502. doi: 10.1016/0042-6822(68)90174-8. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lang D., Bujard H., Wolff B., Russell D. Electron microscopy of size and shape of viral DNA in solutions of different ionic strengths. J Mol Biol. 1967 Jan 28;23(2):163–181. doi: 10.1016/s0022-2836(67)80024-x. [DOI] [PubMed] [Google Scholar]
  19. MACHATTIE L. A., THOMAS C. A., Jr DNA FROM BACTERIOPHAGE LAMBDA: MOLECULAR LENGTH AND CONFORMATION. Science. 1964 May 29;144(3622):1142–1144. doi: 10.1126/science.144.3622.1142. [DOI] [PubMed] [Google Scholar]
  20. MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol. 1962 Jul;5:109–118. doi: 10.1016/s0022-2836(62)80066-7. [DOI] [PubMed] [Google Scholar]
  21. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  23. Signer E. R. Lysogeny: the integration problem. Annu Rev Microbiol. 1968;22:451–488. doi: 10.1146/annurev.mi.22.100168.002315. [DOI] [PubMed] [Google Scholar]
  24. TAYLOR A. L. BACTERIOPHAGE-INDUCED MUTATION IN ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1963 Dec;50:1043–1051. doi: 10.1073/pnas.50.6.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. To C. M., Eisenstark A., Töreci H. Structure of mutator phage Mu1 of Escherichia coli. J Ultrastruct Res. 1966 Mar;14(5):441–448. doi: 10.1016/s0022-5320(66)80074-6. [DOI] [PubMed] [Google Scholar]
  26. Torti F., Barksdale C., Abelson J. Mu-1 bacteriophage DNA. Virology. 1970 Jul;41(3):567–568. doi: 10.1016/0042-6822(70)90179-0. [DOI] [PubMed] [Google Scholar]
  27. Toussaint A. Insertion of phage Mu. 1 within prophage lambda. A new approach for studying the control of the late functions in bacteriophage lambda. Mol Gen Genet. 1969;106(1):89–92. doi: 10.1007/BF00332824. [DOI] [PubMed] [Google Scholar]
  28. VINOGRAD J., BRUNER R., KENT R., WEIGLE J. Band-centrifugation of macromolecules and viruses in self-generating density gradients. Proc Natl Acad Sci U S A. 1963 Jun;49:902–910. doi: 10.1073/pnas.49.6.902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. WILLSON C., PERRIN D., COHN M., JACOB F., MONOD J. NON-INDUCIBLE MUTANTS OF THE REGULATOR GENE IN THE "LACTOSE" SYSTEM OF ESCHERICHIA COLI. J Mol Biol. 1964 Apr;8:582–592. doi: 10.1016/s0022-2836(64)80013-9. [DOI] [PubMed] [Google Scholar]
  30. WYATT G. R. The purine and pyrimidine composition of deoxypentose nucleic acids. Biochem J. 1951 May;48(5):584–590. doi: 10.1042/bj0480584. [DOI] [PMC free article] [PubMed] [Google Scholar]

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