New positive selection system based on the parD (kis/kid) system of the R1 plasmid

Biotechniques. 2000 Apr;28(4):784-8.

Abstract

The use of vectors that are designed to allow positive selection of recombinants facilitates cloning experiments in E. coli. Using kid, a lethal gene of the R1 plasmid parD locus, we generated pKID vectors leading to high selective efficiency of recombinants (greater than 90%). The E. coli bacterial host used to propagate these vectors produces the Kis protein, the natural antagonist of Kid. This new positive-selection system exhibits the same efficiency as the original ccdB-based selection vectors, pKIL (4). We also show that the ccdB and kid systems are independent. This property increases the potential of plasmidic poison-antidote systems for genetic applications and opens the door to a generation of new vectors containing the two selection systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Toxins / genetics*
  • Bacterial Toxins / metabolism
  • Base Sequence
  • Cloning, Molecular / methods
  • DNA-Binding Proteins / genetics*
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Escherichia coli Proteins*
  • Genetic Vectors
  • Molecular Sequence Data
  • R Factors / genetics*
  • R Factors / metabolism

Substances

  • Bacterial Proteins
  • Bacterial Toxins
  • CcdB protein, Plasmid F
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • Kid protein, Bacteria
  • Kis protein, Bacteria
  • ParD protein, E coli
  • ParD protein, Plasmid R1
  • parD protein, Bacteria