Two-dimensional S1 nuclease heteroduplex mapping: Detection of rearrangements in bacterial genomes

T. Yee, M. Inouye

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

A method of two-dimensional S1 nuclease heteroduplex mapping was developed to detect gene rearrangements and repeated sequences in total bacterial chromosomes. To detect DNA rearrangements between two variant bacterial strains, total chromosomal DNA preparations from the two strains are digested with four-base-recognizing restriction enzymes, mixed together, denatured, renatured, and separated on first-dimension polyacrylamide slab gels. Gel strips are cut out and soaked in a buffer containing S1 nuclease, which diffuses into the strips and digests the DNA fragments at single-stranded regions. The digested DNA is then electrophoresed in a second dimension perpendicular to the first dimension. DNA heteroduplexes that were digested by the S1 nuclease are resolved as distinct spots below a bright unresolved band of homoduplex. This report describes testing of this method on a model system consisting of two nearly isogeneic strains of Escherichia coli, and the application of this method in detecting DNA rearrangements associated with phase variation in Myxococcus xanthus.

Original languageEnglish (US)
Pages (from-to)2723-2727
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number9 I
DOIs
StatePublished - 1984
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • General

Fingerprint Dive into the research topics of 'Two-dimensional S1 nuclease heteroduplex mapping: Detection of rearrangements in bacterial genomes'. Together they form a unique fingerprint.

Cite this