RRC ID 33412
Author Snaith MR, Kilby NJ, Murray JA.
Title An Escherichia coli system for assay of F1p site-specific recombination on substrate plasmids.
Journal Gene
Abstract We have developed an Escherichia coli system for testing the behaviour of plasmids carrying target sites for the F1p site-specific recombinase. The E. coli strain BL-FLP is described, which carries a chromosomally integrated bacteriophage T7 RNA polymerase gene expressed from a lac promoter, and harbours the plasmid pMS40.pMS40 has the features: (i) it carries the FLP recombinase gene under the control of a bacteriophage T7 promoter, (ii) it confers kanamycin resistance, and (iii) it uses an R6K origin of replication; these two latter features make it compatible with most conventional cloning vectors. Substrate plasmids carrying F1p-recognition targets (FRT) are transformed into BL-FLP, and the consequences of F1p-mediated recombination can be analysed after subsequent extraction of plasmid DNA. We show that this system is capable of base-perfect F1p-mediated recombination on plasmid substrates. We also present a corrected sequence of the commonly used F1p substrate plasmid, pNEO beta GAL (O'Gorman et al. (1991) Science 251, 1351-1355).
Volume 180(1-2)
Pages 225-7
Published 1996-11-21
PII S0378-1119(96)00449-0
PMID 8973372
MeSH Binding Sites DNA Nucleotidyltransferases / genetics DNA Nucleotidyltransferases / metabolism* Escherichia coli / genetics* Molecular Sequence Data Plasmids / genetics* Plasmids / metabolism Recombination, Genetic*
IF 2.498
Times Cited 6
Prokaryotes E. coli pMS40