In vivo recombineering of bacteriophage lambda by PCR fragments and single-strand oligonucleotides.
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.
Oppenheim
Amos B
AB
Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Rattray
Alison J
AJ
Bubunenko
Mikhail
M
Thomason
Lynn C
LC
Court
Donald L
DL
D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D017353:Gene Deletion; D005818:Genetic Engineering / Q000379:methods; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics*; D017354:Point Mutation; D011995:Recombination, Genetic
D009841:Oligonucleotides