Combinatorial screening of mixed populations of organisms
AssigneeDiversa Corporation
InventorJay M. Short
Life of this patent
Estimating the term…
Abstract
Provided is a method of screening gene libraries derived from a mixed population of organisms for a bioactivity or biomolecule of interest. The mixed population of organisms can be a cultured population or an uncultured population from, for example, the environment. Also provided are methods of screening isolates or enriched populations of organisms, which isolates include a population that is spatially, temporally, or hierarchical, for example, of a particular species, genus, family, or class of organisms. Identified clones containing a biomolecule or bioactivity of interest can be further variegated or the DNA contained in the clone can be variegated to create novel biomolecules or bioactivities of interest.
Claims30
A method for identifying a protein having an activity of interest, comprising:
- (a) incubating nucleic acids obtained directly without selection from a mixed population of organisms from an environmental source, said nucleic acid has at least a portion of a nucleic acid sequence encoding a molecule of interest, with at least one oligonucleotide probe labeled with a detectable label under such conditions and such time to allow interaction of complementary sequences;
- (b) identifying nucleic acid sequences having a complement to the oligonucleotide probe using an analyzer that detects the detectable label;
- (c) generating a library from the identified nucleic acid sequences;
- (d) screening the library for a specified activity;
- (e) mutating a nucleic acid sequence contained in a clone from the library having the specified activity;
- and
- (d) comparing the activity of an expression product of the clone from (e) following mutation with the specified activity of an expression product of the clone without mutation, wherein a difference in the activity is indicative of an effect of introducing at least one sequence mutation, thereby identifying a protein having an activity of interest.
Dependencies are inferred from the claim text.