Simple assay for quantitation of Tn5 inversion events in Escherichia coli and use of the assay in determination of plasmid copy number.
In this report, we describe a simple method for measuring the frequency of sequence inversion in the transposable element Tn5 as a result of recombination across its duplicated IS50 elements. The structure of Tn5 was manipulated so that the neomycin phosphotransferase gene of the transposon would be expressed only if a sequence inversion event occurred. This highly sensitive assay also served as the basis for a novel means of estimating plasmid copy number.
Weber
P C
PC
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109.
Levine
M
M
Glorioso
J C
JC
D007446:Chromosome Inversion; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D010957:Plasmids; D011938:Rec A Recombinases / Q000502:physiology; D011995:Recombination, Genetic; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D011938:Rec A Recombinases