US6054267A
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US
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Method for screening for enzyme activity
Assignee: BP Corp North America Inc
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Filing: 1996-08-02
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Granted: 2000-04-25
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Expires: 2016-08-02
Abstract
Abstract Disclosed is a process for identifying clones having a specified enzyme activity by screening for the specified enzyme activity in a library of clones prepared by (i) selectively isolating target DNA from DNA derived from at least one microorganism, by use of at least one probe DNA comprising at least a portion of a DNA sequence encoding an enzyme having the specified enzyme activity; and (ii) transforming a host with isolated target DNA to produce a library of clones which are screened for the specified enzyme activity.
Inventors
- 👤 Jay M. Short
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 21:15
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/16; C12N15/00; C12N15/09; C12N15/10; C12N15/102; C12N15/1034; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2434; C12Q; C12Q1/00; C12Q1/68; C12Q1/6811; C12Y; C12Y301/00; C12Y301/03; C12Y301/03001; C12Y302/00; C12Y302/01; C12Y302/01021
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
84 words
A method for enriching for DNA sequences containing at least a partial coding region for at least one specified enzyme activity in a DNA sample comprising selecting and recovering a mixture of target DNA from a mixture of organisms, by use of a mixture of DNA probes comprising at least a portion of a DNA sequence encoding at least one enzyme having a specified enzyme activity and transforming host cells with recovered target DNA to produce an expression library of a plurality of clones.
Claim 2
Independent Claim
84 words
A method for enriching for DNA sequences containing at least a partial coding region for at least one specified enzyme activity in a DNA sample comprising selecting and recovering a mixture of target DNA from a mixture of organisms, by use of a mixture of DNA probes comprising at least a portion of a DNA sequence encoding at least one enzyme having a specified enzyme activity and transforming host cells with recovered target DNA to produce an expression library of a plurality of clones.
Claim 3
Dependent Claim
10 words
The method of claim 1, wherein the organisms are microorganisms.
Claim 4
Dependent Claim
11 words
The method of claim 2, wherein the microorganisms are uncultured microorganisms.
Claim 5
Dependent Claim
16 words
The method of claim 1, further comprising screening the expression library for the specified enzyme activity.
Claim 6
Dependent Claim
67 words
The method of claim 1, wherein the target DNA obtained from the DNA population is selected by: a) converting double stranded DNA into single stranded DNA; b) recovering from the converted single stranded DNA, single stranded target DNA which hybridizes to probe DNA; c) converting recovered single stranded target DNA to double stranded DNA; and d) transforming a host cell with the double stranded DNA of c).
Claim 7
Dependent Claim
31 words
The method of claim 5, wherein the probe is directly or indirectly bound to a solid phase by which it is separated from single stranded DNA which is not so hybridized.
Claim 8
Dependent Claim
31 words
The method of claim 6, which further comprises: a) releasing single stranded DNA from said probe; and b) amplifying the released single stranded DNA prior to converting to double stranded DNA.
Claim 9
Dependent Claim
13 words
The method of claim 1, wherein said target DNA is gene cluster DNA.
Claim 10
Dependent Claim
15 words
The method of claim 3, wherein the uncultured microorganisms are derived from an environmental sample.
Claim 11
Dependent Claim
32 words
The method of claim 3, wherein the uncultured microorganisms comprise a mixture of terrestrial microorganisms or marine microorganisms or airborne microorganisms, or a mixture of terrestrial microorganisms, marine microorganisms and airborne microorganisms.
Claim 12
Dependent Claim
27 words
The method of claim 1, wherein the clones comprise a construct selected from the group consisting of phage, plasmids, phagemids, cosmids, phosmids, viral vectors, and artificial chromosomes.
Claim 13
Dependent Claim
21 words
The method of claim 1, wherein the target DNA comprises one or more operons, or portions thereof, of the DNA population.
Claim 14
Dependent Claim
18 words
The method of claim 12, wherein the operon or portions thereof encodes a complete or partial metabolic pathway.
Claim 15
Dependent Claim
11 words
The method of claim 3, wherein the uncultured microorganisms comprise extremophiles.
Claim 16
Dependent Claim
21 words
The method of claim 14, wherein the extremophiles are selected from the group consisting of thermophiles, hyperthermophiles, psychrophiles, barophiles, and psychrotrophs.
Claim 17
Dependent Claim
26 words
The method of claim 4, wherein the host cell is selected from the group consisting of a bacterium, fungus, plant cell, insect cell and animal cell.
Claim 18
Dependent Claim
12 words
The method of claim 1, wherein the target DNA encodes a protein.
Claim 19
Dependent Claim
11 words
The method of claim 17, wherein the protein is an enzyme.
Claim 20
Dependent Claim
22 words
The method of claim 18 wherein the enzyme is selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases.
Claim 21
Independent Claim
52 words
A method for making a gene library, comprising randomly concatenating cDNA or genomic DNA fragments obtained from one or more species of uncultured organisms, and ligating the concatenated DNA fragments to a DNA vector to generate a library of constructs in which genes are contained in the cDNA or genomic DNA fragments.
Claim 22
Independent Claim
52 words
A method for making a gene library, comprising randomly concatenating cDNA or genomic DNA fragments obtained from one or more species of uncultured organisms, and ligating the concatenated DNA fragments to a DNA vector to generate a library of constructs in which genes are contained in the cDNA or genomic DNA fragments.
Claim 23
Dependent Claim
27 words
The method of claim 20, wherein the uncultured organisms comprise a mixture of terrestrial microorganisms or marine microorganisms, or a mixture of terrestrial microorganisms and marine microorganisms.
Claim 24
Dependent Claim
15 words
The method of claim 20, wherein the uncultured organisms are derived from an environmental sample.
Claim 25
Dependent Claim
23 words
The method of claim 20, wherein the cDNA or genomic fragments comprise at least an operon, or portions thereof, of the donor microorganisms.
Claim 26
Dependent Claim
15 words
The method of claim 23, wherein the operon encodes a complete or partial metabolic pathway.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2003-04-25 | 2003-10-25 | 2004-04-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-04-25 | 2007-10-25 | 2008-04-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-04-25 | 2011-10-25 | 2012-04-25 | $7700.00 | Window Closed / Lapsed |