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genetic footprinting

An approach to the identification of the function of a large number of putative genes of a micro-organism, such as may have been uncovered by genome sequencing. A transposable element is inserted into a large number of sites in the genome of the micro-organism, and the mutagenized population is then grown under a wide variety of conditions that may suggest a gene's function. After many population doublings under each set of conditions, the micro-organisms' DNA is extracted and used as a resource for analysis of as many of the putative genes as is desired. PCR primers are constructed to hybridize with the transposable element and with a putative gene. PCR amplification using the primer pair will produce a spectrum of bands on separation by polyacrylamide-gel electrophoresis, one from each mutation that can grow under the set of conditions, i.e. a genetic footprint of the gene. A comparison of the gene's footprints under permissive and restrictive conditions will indicate, by the absence of bands in the latter footprint, those growth conditions that require the function of the gene. Smith, V., Botstein, D. and Brown, P.O. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 6479-6483

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