Open AccessGene replacement and retrieval with recombinant M13mp bacteriophages
Author(s) -
Paul H. Blum,
Donald L. Holzschu,
Hoi Shan Kwan,
Daniel L. Riggs,
S W Artz
Publication year - 1989
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.171.1.538-546.1989
Subject(s) - biology , lysogen , operon , genetics , escherichia coli , gene , recombinant dna , allele , salmonella , bacteriophage , bacteria
We have developed an allele exchange system for shuttling sequences of DNA to and from their original chromosomal loci. Cloned segments of the histidine operon of Salmonella typhimurium and the lactose operon of Escherichia coli served as target sequences and were used to develop the system. Replacement and retrieval of target sequences used the phage M13mp vectors and proceeded through an M13 lysogen intermediate. The intermediates and products of allele exchange were characterized by genetic and hybridization analyses. Several unique properties of M13 lysogens were exploited to devise positive selections to detect integration and excision. These positive selections were used to manipulate phenotypically silent alleles.