Biosensor materials and methods

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor

Reexamination Certificate

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C435S320100, C435S476000, C536S023100, C536S024100

Reexamination Certificate

active

06849442

ABSTRACT:
Disclosed are methods for generating mycolic acid bacterial biosensors for particular analytes (especially industrial pollutants) by the use of innovative methods for isolating DNA encoding an inducible promoter which is induced in response to the specific analyte (and/or associated operon proteins), the methods generally comprising the steps of: (a) culturing a source of mycolic acid bacteria in a selective medium containing said specific analyte and being selective for oligotriphic bacteria; (b) identifying mycolic acid bacteria capable of subsisting on said medium, especially those which do not display catabolic repression; (c) extracting DNA from said mycolic acid bacteria; (d) incorporating said DNA into vectors, such as various shuttle vectors; (e) cloning said vector into a suitable host cell (which may beE. colistrain carrying one or more of the mcrABC mrr hsdSRM recA and recO mutations); (f) screening that host cell (or a second host cell which is preferably acorynebacterium) for said inducible promoter. The methods are exemplified by the isolation of theR. corallinaohp operon.

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