In vivo recombineering of bacteriophage lambda by PCR fragments and single-strand oligonucleotides

Virology. 2004 Feb 20;319(2):185-9. doi: 10.1016/j.virol.2003.11.007.

Abstract

We demonstrate that the bacteriophage lambda Red functions efficiently recombine linear DNA or single-strand oligonucleotides (ss-oligos) into bacteriophage lambda to create specific changes in the viral genome. Point mutations, deletions, and gene replacements have been created. While recombineering with oligonucleotides, we encountered other mutations accompanying the desired point mutational change. DNA sequence analysis suggests that these unwanted mutations are mainly frameshift deletions introduced during oligonucleotide synthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriophage lambda / genetics*
  • Base Sequence
  • Gene Deletion
  • Genetic Engineering / methods
  • Molecular Sequence Data
  • Oligonucleotides / genetics*
  • Point Mutation
  • Recombination, Genetic

Substances

  • Oligonucleotides