Phage lambda and plasmid expression vectors with multiple cloning sites and lacZ alpha-complementation

Gene. 1986;45(1):95-9. doi: 10.1016/0378-1119(86)90136-8.

Abstract

Two new lambda vectors were constructed which permit cloning of genes that are potentially lethal if cloned in analogous plasmid vectors. lambda DL10 and lambda DL11 contain the alpha-complementing fragment of lacZ and multiple cloning sites found in the polylinker region of M13mp10 and M13mp11, respectively. DNA cloned into the unique cloning sites of these vectors can be detected by inactivation of alpha-complementation. These lambda vectors provide a lac promoter for expression of foreign genes as well as the ability to make fusion proteins. Two plasmid expression vectors, pPR110 and pPR111, were constructed from lambda DL10 and lambda DL11 respectively, and pCQV2. These plasmids, which express lacZ alpha-complementing activity from the lambda PR promoter, contain multiple cloning sites immediately downstream of the PR promoter. They allow cloning of genes under the control of the PR promoter and the plasmid-encoded thermosensitive (cI857) repressor, and allow easy detection of inserted fragments by inactivation of alpha-complementation.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacteriophage lambda / genetics*
  • Cloning, Molecular / methods
  • Coliphages / genetics
  • Genes, Lethal
  • Genes, Viral
  • Genetic Complementation Test
  • Genetic Vectors*
  • Lac Operon*
  • Plasmids*
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / genetics
  • beta-Galactosidase / genetics

Substances

  • Bacterial Proteins
  • Recombinant Fusion Proteins
  • beta-Galactosidase