Nucleic Acids pp 245-250 | Cite as

In Vitro Packaging of DNA

  • J. W. Dale
  • P. J. Greenaway
Part of the Methods in Molecular Biology book series (MIMB, volume 2)

Abstract

In the normal growth cycle of bacteriophage lambda, the proteins that ultimately form the head of the phage particle are assembled into an empty precursor of the head (prehead); the phage DNA is replicated separately and then inserted into the empty head particles—a process known as packaging. A number of phage gene products play an important role in this process. Amongst these are:
  1. (i)

    The E protein, which is the major component of the phage head; mutants that are defective in this gene are unable to assemble the preheads, and therefore accumulate the other, unassembled, components of the phage particle as well as the other proteins involved in packaging.

     
  2. (ii)

    The D and A proteins, which are involved in the packaging process itself. Mutants that are defective in these genes are able to produce the preheads, but will not package DNA. This results in the accumulation of empty preheads.

     

Keywords

Sucrose Agar EDTA Foam MgCl2 

References

  1. 1.
    Collins, J. and Hohn, E. (1978) Cosmids: A type of plasmid gene cloning vector that is packageable in vitro in bacterio-phage heads. Proc. Natl. Acad. Sci. USA 75, 4242–4246.PubMedCrossRefGoogle Scholar
  2. 2.
    Enquist, L., and Sternberg, N. (1979) In vitro packaging of lambda Dam vectors and their use in cloning DNA fragments. Meth. Enzymology 68, 281–298.CrossRefGoogle Scholar
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    Hohn, B. (1979) In vitro packaging of lambda and cosmid DNA. Meth. Enzymology 68, 299–309.CrossRefGoogle Scholar
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    Hohn, B., and Murray, K. (1977) Packaging recombinant DNA molecules into bacteriophage particles in vitro. Proc. Natl. Acad. Sci. USA 74, 3259–3263.PubMedCrossRefGoogle Scholar
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    Sternberg, N., Tiemeier, D., and Enquist, L. (1977) In vitro packaging of a lambda dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. Gene 1, 255–280.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc. 1984

Authors and Affiliations

  • J. W. Dale
    • 1
    • 2
  • P. J. Greenaway
    • 1
    • 2
  1. 1.Department of MicrobiologyUniversity of SurreyGuildford
  2. 2.Molecular Genetics LaboratoryPHLS Centre for Applied Microbiology and Research, Porton DownSalisburyUK

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