Lambda as a Cloning Vector

Karen Lech1, Roger Brent1, Nina Irwin2

1 Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, 2 Harvard University, Cambridge, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 1.10
DOI:  10.1002/0471142727.mb0110s00
Online Posting Date:  May, 2001
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Advantages of lambda as a cloning vector are discussed along with considerations for the insert DNA (i.e., size, spi and hfl state). Maps of lambda‐derived cloning vectors are provided in addition to a discussion and map of a cosmid (a lambda‐derived plasmid vector).

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Table of Contents

  • Advantages of Using Lambda
  • Selections for Inserted DNA
  • Maps of Lambda‐Derived Cloning Vectors
  • The Cosmid, a Useful Lambda‐Derived Plasmid Vector
  • Figures
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Literature Cited

Literature Cited
   Banuett, F., Hoyt, A.M., McFarlane, L., Echols, H., and Herskowitz, I. 1986. hflB, a new Escherichia coli locus regulating lysogeny and the level of bacteriophage lambda cII protein. J. Mol. Biol. 187:213‐224.
   Blattner, F.R., Williams, B.G., Blechl, A.E., Denniston‐Thompson, K., Faber, H.E., Furlong, L.‐A., Grunwald, D.J., Kiefer, D.O., Moore, D.D., Schumm, J.W., Sheldon, E.O., and Smithies, O. 1977. Charon phages: Safer derivatives of bacteriophage lambda for DNA cloning Science 196:161‐169.
   deWet, J.R., Daniels, D.L., Schroeder, J.L., Williams, B.G., Denniston‐Thompson, K., Moore, D.D., and Blattner, F.R. 1980. Restriction maps for twenty‐one Charon vector phages. J. Virol. 33:401‐410.
   Dunn, I.S. Blattner, F.R. 1987. Charon 36 and 40: Multi‐enzyme, high capacity, recombination deficient replacement vectors with polylinkers and polystuffers. Nucl. Acids Res. 15:2677‐2698.
   Frischauf, A.‐M., Lehrach, H., Polstka, A., and Murray, N.M. 1983. Lambda replacement vectors carrying polylinker sequences. J. Mol. Biol. 170:827‐842.
   Gibson, T.J., Coulson, A.R., Sulston, J.E., and Little, P.F.R. 1987. Lorist 2, a cosmid with transcriptional terminators insulating vector genes from interference by promoters within the insert: Effect on DNA yield and cloned insert frequency. Gene 53:275‐281.
   Hendrix, R., Roberts, J., Stahl, F., and Weisberg, R. 1983. Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
   Hoyt, M.A., Knight, D.M., Das, A., Miller, H.I., and Echols, H. 1982. Control of phage lambda development by stability and synthesis of cII protein: Role of the viral cIII and host hflA, himA, and himD genes. Cell 31:565‐573.
   Huynh, T.V., Young, R.A., and Davis, R.W. 1984. Constructing and screening cDNA libraries in λgt10 and λgt11. In DNA Cloning Techniques: A Practical Approach (D. Glover, ed.) pp. 49‐78. IRL Press, Oxford.
   Ish‐Horowitz, D. Burke, J.F. 1981. Rapid and efficient cosmid cloning. Nucl. Acids Res. 9:2989‐2998.
   Karn, J., Matthes, H.W.P., Gait, M.T., and Brenner, S. 1984. A new selection cloning vector, λ2001, with sites for XbaI, BamHI, HindIII, EcoRI, SstI, and XhoI. Gene 32:217‐224.
   Williams, B.G. Blattner, F.R. 1979. Construction and characterization of the hybrid bacteriophage lambda Charon vectors for DNA cloning. J. Virol. 29:555‐575.
   Young, R.A. Davis, R.W. 1983. Efficient isolation of genes by using antibody probes. Proc. Natl. Acad. Sci. U.S.A. 80:1194‐1198.
   Zissler, J., Signer, E., and Schaefer, F. 1971. The role of recombination in growth of bacteriophage lambda. In The Bacteriophage Lambda (A.D. Hershey, ed.) pp. 455‐475. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Key Reference
   Murray, N.E. 1983. Phage Lambda and Molecular Cloning. In Lambda II, (R.W. Hendrix, J. Roberts, F. Stahl, and R. Weisberg, eds.) pp. 395‐432. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
  Provides a thorough introduction to the biology of cloning vectors in common use before 1983.
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