A Small Cosmid for Efficient Cloning of Large DNA Fragments

Gene. 1980 Nov;11(3-4):291-8. doi: 10.1016/0378-1119(80)90069-4.

Abstract

The production and use of the 6 kb cosmid pHC79, a derivative of pBR322, is described. It can be used for cloning of fragments cleaved by EcoRI, ClaI, BamHI (also BglII, BclI, Sau3A and MboI), SalI (also XhoI and AvaI), EcaI and PstI. Hybrid cosmids containing inserts in the size range of 40 kb are packaged in vitro and transduced with an efficiency of 5 X 10(4) - 5 X 10(5) clones/microgram of insert DNA. Prefractionation of the DNA fragments to be cloned into 40 kb sized fragments ensures the cloning of contiguous stretches of DNA. Proteins produced in vitro by the cosmid pHC79 are identical to the ones produced by its pBR322 parent.

MeSH terms

  • Bacteriophage lambda / genetics
  • Cloning, Molecular / methods*
  • DNA Restriction Enzymes
  • Escherichia coli / genetics
  • Genetic Vectors*
  • Molecular Weight
  • Plasmids
  • Proteins / genetics

Substances

  • Proteins
  • DNA Restriction Enzymes