Evidence for a prokaryotic insertion-sequence contamination in eukaryotic sequences registered in different databases.
An insertion-sequence of prokaryotic origin was detected in a genomic clone obtained from a Phaseolus vulgaris bacterial artificial chromosome (BAC) library. This BAC clone, characterized as part of a contig constructed near a virus resistance gene, exhibited restriction fragment length polymorphism with an overlapping clone of the contig. Restriction analysis of DNA obtained from individual colonies of the stock culture indicated the presence of a mixed population of wild-type and insertional mutants. Sequence analysis of both members of the population revealed the presence of IS 10R, an insertion-sequence from Escherichia coli. A BLAST search for IS 10-like sequences detected unexpected homologies with a large number of eukaryotic sequences from Homo sapiens, Arabidopsis thaliana, Drosophila melanogasterand Caenorhabditis elegans. Southern analysis of a random sample of BAC clones failed to detect IS 10 in the BAC DNA. However, prolonged sub-culturing of a set of 15 clones resulted in transposition into the BAC DNA. Eventually, all cultures acquired a 2.3-kb fragment that hybridized strongly with IS 10. Sequence analysis revealed the presence of a preferred site for transposition in the BAC vector. These results indicate that a large number, if not all, of the BAC libraries from different organisms are contaminated with IS 10R. The source of this element has been identified as the DH10B strain of E. coli used as the host for BAC libraries.
Astua-Monge
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Department of Horticultural Sciences, and Plant Molecular and Cellular Biology Graduate Program, University of Florida, Gainesville, FL 32611-0690, USA.
Lyznik
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Jones
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Mackenzie
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Vallejos
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