Compartmentalization method for screening microorganisms

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism

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435 29, 435 14, 435 15, 435 25, 435 23, 435 24, 435 28, 4352992, 4353001, 4353042, C12Q 104, C12Q 102, C12Q 154

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060015861

ABSTRACT:
A method is provided for screening microorganisms having a selectable characteristic comprising: (a) providing a growth chamber capable of forming a plurality of discrete compartments, said compartments comprising a barrier which inhibits the diffusion of metabolites or nutrients between said compartments within said growth chamber; (b) preparing a growth medium which, by its lack of or addition of one or more metabolites or nutrients, preferentially facilitates growth of microorganisms exhibiting said selectable characteristic from microorganisms which do not exhibit said selectable characteristic; (c) inoculating said growth medium with a plurality of microorganisms, a portion of which comprise said selectable characteristic; (d) placing said inoculated growth medium into said growth chamber so as to randomly disperse said microorganisms among discrete compartments in said growth chamber; and (e) incubating said growth chamber for a suitable time and under suitable conditions to obtain a detectable disparity of growth between microorganisms having said selectable characteristic and microorganisms lacking said selectable characteristic.

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Joyce, "Directed Molecular Evolution," Scientific American, Dec. 1992, pp. 90-97.
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Moore et al. "Directed Evolution of a Para-Nitrobenzyl Esterase for Aqueous-Organic Solvents," Nature Biotechnology, 14:458-467, Apr. 1996.
Osuna et al. "Microbial Systems and Directed Evolution of Protein Activities," Critical Reviews in Microbiology, 20(2):107-116, 1994.

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