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Series GSE7695

Query DataSets for GSE7695

Status

Public on Sep 30, 2007

Title

Caveat Mutator

Organism

Escherichia coli

Experiment type

Expression profiling by array
Genome variation profiling by array

Summary

The use of whole genome microarrays for monitoring mutagenised or otherwise engineered genetic derivatives is a potentially powerful tool for checking genomic integrity. Using comparative genomic hybridization of a number of unrelated, directed deletion mutants in Escherichia coli K-12 MG1655 we identified unintended secondary genomic deletions in the flhDC region in deltafnr, deltacrp, and deltacreB mutants. These deletions were confirmed by PCR and phenotypic tests. Our findings show that non-motile progeny are found in some populations of MG1655 directed deletion mutants, and studies on the effects of gene knock-outs should be viewed with caution when the mutants have not been screened for the presence of secondary deletions, or confirmed by other methods.
Keywords: comparative genomic hybridisation

Overall design

We used the CGH method to genetically characterize a series of regulatory gene deletion mutants we had made in MG1655 using the lamda-Red method of Datsenko and Wanner. A number of strains were tested using CGH, and each strain was tested only once. Genomic DNA isolated from wt MG1655 was used as a reference in all hybridisations.

Contributor(s)

Hobman JL, Patel M, Hidalgo-Arroyo AG, Cariss JL, Avison MB, Penn CW, Constantinidou C

Citation(s)

17921306

Submission date

May 03, 2007

Last update date

Apr 26, 2012

Contact name

Chrystala Constantinidou

E-mail(s)

C.Constantinidou@bham.ac.uk