초록 ▼

Disclosed is a process for identifying clones having a specified activity of interest, which process comprises (i) generating one or more expression libraries derived from nucleic acid directly isolated from the environment; and (ii) screening said libraries utilizing an assay system. More particula

Disclosed is a process for identifying clones having a specified activity of interest, which process comprises (i) generating one or more expression libraries derived from nucleic acid directly isolated from the environment; and (ii) screening said libraries utilizing an assay system. More particularly, this is a process for identifying clones having a specified activity of interest by (i) generating one or more expression libraries derived from nucleic acid directly or indirectly isolated from the environment; (ii) exposing said libraries to a particular substrate or substrates of interest; and (iii) screening said exposed libraries utilizing a fluorescence activated cell sorter to identify clones which react with the substrate or substrates. Also provided is a process for identifying clones having a specified activity of interest by (i) generating one or more expression libraries derived from nucleic acid directly or indirectly isolated from the environment; and (ii) screening said exposed libraries utilizing an assay requiring a binding event or the covalent modification of a target, and a fluorescence activated cell sorter to identify positive clones.

대표청구항 ▼

[ What is claimed is:] [1.] A method for identifying a desired activity encoded by a genomic DNA population comprising:(a) obtaining a single-stranded genomic DNA population;(b) contacting the single-stranded DNA population of (a) with a DNA probe bound to a ligand under conditions and for sufficien

[ What is claimed is:] [1.] A method for identifying a desired activity encoded by a genomic DNA population comprising:(a) obtaining a single-stranded genomic DNA population;(b) contacting the single-stranded DNA population of (a) with a DNA probe bound to a ligand under conditions and for sufficient time to allow hybridization and to produce a double-stranded complex of probe and members of the genomic DNA population which hybridize thereto;(c) contacting the double-stranded complex of (b) with a solid phase specific binding partner for said ligand so as to produce a solid phase complex;(d) separating the solid phase complex from the single-stranded DNA population of (b);(e) releasing from the probe the members of the genomic population which had bound to the solid phase bound probe;(f) forming double-stranded DNA from the members of the genomic population of (e);(g) introducing the double-stranded DNA of (f) into a suitable host cell to produce an expression library containing a plurality of clones containing and expressing the selected DNA;(h) producing a cell-free extract of the expression library,(i) combining the expression library extract with a host cell-free protein extract from a metabolically rich host organism to form an extract mixture, and(j) screening the extract mixture for a desired activity resulting from the combining of the cell-free extracts.