Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Dec;80(24):7456–7460. doi: 10.1073/pnas.80.24.7456

DNA sequences necessary for packaging of bacteriophage lambda DNA.

B Hohn
PMCID: PMC389970  PMID: 6324174

Abstract

The extent of DNA flanking the "cohered cohesive end" site of bacteriophage lambda DNA, which is required for packaging, was determined by using defined DNA fragments and a cosmid in vivo packaging assay. From the right end of lambda DNA a 20- to 36-base-pair stretch extending from the center of the cohered cohesive ends is shown to be required, whereas the packaging efficiency of cosmids extending to 70 base pairs into the left lambda arm is reduced to 10% (compared to a fragment extending until about 80 base pairs). A 60-base-pair stretch of the left arm leaves an efficiency of only 1%. The segment thus delineated, by the nature of the assay, is both necessary and sufficient for the binding of packaging proteins to the DNA, the packaging of DNA itself, the DNA cleavage, and successful injection of the DNA into a bacterial host. By contrast, in vitro packaging of restriction fragments of mature lambda DNA directly demonstrated the selectivity of the packaging proteins for the fragment originating from the left end of the DNA. The results of the two complementary experiments are discussed in terms of the various steps before, during, and after packaging for which different sequences flanking and including the cohered cohesive ends might be required.

Full text

PDF
7456

Images in this article

7459Image
on p.7459

Selected References

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

  1. Collins J., Hohn B. Cosmids: a type of plasmid gene-cloning vector that is packageable in vitro in bacteriophage lambda heads. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4242–4246. doi: 10.1073/pnas.75.9.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Emmons S. W. Bacteriophage lambda derivatives carrying two copies of the cohesive end site. J Mol Biol. 1974 Mar 15;83(4):511–525. doi: 10.1016/0022-2836(74)90511-7. [DOI] [PubMed] [Google Scholar]
  3. Feiss M., Campbell A. Duplication of the bacteriophage lambda cohesive end site: genetic studies. J Mol Biol. 1974 Mar 15;83(4):527–540. doi: 10.1016/0022-2836(74)90512-9. [DOI] [PubMed] [Google Scholar]
  4. Feiss M., Fisher R. A., Crayton M. A., Egner C. Packaging of the bacteriophage lambda chromosome: effect of chromosome length. Virology. 1977 Mar;77(1):281–293. doi: 10.1016/0042-6822(77)90425-1. [DOI] [PubMed] [Google Scholar]
  5. Feiss M., Fisher R. A., Siegele D. A., Nichols B. P., Donelson J. E. Packaging of the bacteriophage lambda chromosome: a role for base sequences outside cos. Virology. 1979 Jan 15;92(1):56–67. doi: 10.1016/0042-6822(79)90214-9. [DOI] [PubMed] [Google Scholar]
  6. Feiss M., Kobayashi I., Widner W. Separate sites for binding and nicking of bacteriophage lambda DNA by terminase. Proc Natl Acad Sci U S A. 1983 Feb;80(4):955–959. doi: 10.1073/pnas.80.4.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feiss M., Siegele D. A., Rudolph C. F., Frackman S. Cosmid DNA packaging in vivo. Gene. 1982 Feb;17(2):123–130. doi: 10.1016/0378-1119(82)90064-6. [DOI] [PubMed] [Google Scholar]
  8. Feiss M., Widner W. Bacteriophage lambda DNA packaging: scanning for the terminal cohesive end site during packaging. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3498–3502. doi: 10.1073/pnas.79.11.3498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feiss M., Widner W., Miller G., Johnson G., Christiansen S. Structure of the bacteriophage lambda cohesive end site: location of the sites of terminase binding (cosB) and nicking (cosN). Gene. 1983 Oct;24(2-3):207–218. doi: 10.1016/0378-1119(83)90081-1. [DOI] [PubMed] [Google Scholar]
  10. Hohn B. DNA as substrate for packaging into bacteriophage lambda, in vitro. J Mol Biol. 1975 Oct 15;98(1):93–106. doi: 10.1016/s0022-2836(75)80103-3. [DOI] [PubMed] [Google Scholar]
  11. Hohn B., Wurtz M., Klein B., Lustig A., Hohn T. Phage lambda DNA packaging, in vitro. J Supramol Struct. 1974;2(2-4):302–317. doi: 10.1002/jss.400020220. [DOI] [PubMed] [Google Scholar]
  12. Hohn T., Katsura I. Structure and assembly of bacteriophage lambda. Curr Top Microbiol Immunol. 1977;78:69–110. doi: 10.1007/978-3-642-66800-5_3. [DOI] [PubMed] [Google Scholar]
  13. Hohn T., Wurtz M., Hohn B. Capsid transformation during packaging of bacteriophage lambdaDNA. Philos Trans R Soc Lond B Biol Sci. 1976 Nov 30;276(943):51–61. doi: 10.1098/rstb.1976.0097. [DOI] [PubMed] [Google Scholar]
  14. Meyerowitz E. M., Guild G. M., Prestidge L. S., Hogness D. S. A new high-capacity cosmid vector and its use. Gene. 1980 Nov;11(3-4):271–282. doi: 10.1016/0378-1119(80)90067-0. [DOI] [PubMed] [Google Scholar]
  15. Miwa T., Matsubara K. Formation of oligomeric structures from plasmid DNA carrying cos lambda that is packaged into bacteriophage lambda heads. J Bacteriol. 1983 Jan;153(1):100–108. doi: 10.1128/jb.153.1.100-108.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miwa T., Matsubara K. Identification of sequences necessary for packaging DNA into lambda phage heads. Gene. 1982 Dec;20(2):267–279. doi: 10.1016/0378-1119(82)90045-2. [DOI] [PubMed] [Google Scholar]
  17. Miwa T., Matsubara K. Lambda phage DNA sequences affecting the packaging process. Gene. 1983 Oct;24(2-3):199–206. doi: 10.1016/0378-1119(83)90080-x. [DOI] [PubMed] [Google Scholar]
  18. Nichols B. P., Donelson J. E. 178-Nucleotide sequence surrounding the cos site of bacteriophage lambda DNA. J Virol. 1978 May;26(2):429–434. doi: 10.1128/jvi.26.2.429-434.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sanger F., Coulson A. R., Hong G. F., Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage lambda DNA. J Mol Biol. 1982 Dec 25;162(4):729–773. doi: 10.1016/0022-2836(82)90546-0. [DOI] [PubMed] [Google Scholar]
  20. Serwer P. Buoyant density sedimentation of macromolecules in sodium iothalamate density gradients. J Mol Biol. 1975 Mar 5;92(3):433–448. doi: 10.1016/0022-2836(75)90290-9. [DOI] [PubMed] [Google Scholar]
  21. Thomas R. Control of development in temperate bacteriophages. 3. Which prophage genes are and which are not trans-activable in the presence of immunity? J Mol Biol. 1970 Apr 28;49(2):393–404. doi: 10.1016/0022-2836(70)90252-4. [DOI] [PubMed] [Google Scholar]
  22. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES