CA2227342A1 - Screening methods for enzymes and enzyme kits - Google Patents
Screening methods for enzymes and enzyme kitsInfo
- Publication number
- CA2227342A1 CA2227342A1 CA002227342A CA2227342A CA2227342A1 CA 2227342 A1 CA2227342 A1 CA 2227342A1 CA 002227342 A CA002227342 A CA 002227342A CA 2227342 A CA2227342 A CA 2227342A CA 2227342 A1 CA2227342 A1 CA 2227342A1
- Authority
- CA
- Canada
- Prior art keywords
- dna
- clones
- screening
- enzyme
- library
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1086—Preparation or screening of expression libraries, e.g. reporter assays
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1079—Screening libraries by altering the phenotype or phenotypic trait of the host
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
Abstract
Recombinant enzyme libraries and kits where a plurality of enzymes are each characterized by different physical and/or chemical characteristics and classified by common characteristics. The characteristics are determined by screening of recombinant enzymes expressed by a DNA library produced from various microorganisms. Also disclosed is a process for identifying clones of a recombinant library which express a protein with a desired activity by screening a library of expression clones randomly produced from DNA of at least one microorganism, said screening being effected on expression products of said clones to thereby identify clones which express a protein with a desired activity. Also disclosed is a process of screening clones having DNA from an uncultivated microorganism for a specified protein activity by screening for a specified protein activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein activity.
Claims (13)
1. An enzyme kit, comprising:
a package said package containing at least three different recombinant enzymes, each of said at least three different recombinant enzymes having at least two enzyme characteristics in common, one of said at least two common enzyme characteristics being a chemical characteristic and the other of said at least two common enzyme characteristics being a physical characteristic.
a package said package containing at least three different recombinant enzymes, each of said at least three different recombinant enzymes having at least two enzyme characteristics in common, one of said at least two common enzyme characteristics being a chemical characteristic and the other of said at least two common enzyme characteristics being a physical characteristic.
2. A process for obtaining a recombinant enzyme library, comprising:
screening recombinant proteins produced by a plurality of expression clones, each of which contains a polynucleotide derived from a microorganism, said screening being effected to determine the clones which produce recombinant enzymes and to determine a plurality of different enzyme characteristics for the recombinant enzymes. and classifying the recombinant enzymes by said enzyme characteristics.
screening recombinant proteins produced by a plurality of expression clones, each of which contains a polynucleotide derived from a microorganism, said screening being effected to determine the clones which produce recombinant enzymes and to determine a plurality of different enzyme characteristics for the recombinant enzymes. and classifying the recombinant enzymes by said enzyme characteristics.
3. The process of Claim 2 wherein said screening includes screening said recombinant enzymes for a chemical characteristic, and rescreening recombinant enzymes having said chemical characteristic for a second chemical characteristic.
4. A process for producing an enzyme screening kit. comprising:
placing in a package at least three different selected enzymes, said enzymes being selected from a recombinant enzyme library having a plurality of differentenzymes having different enzyme characteristics, said enzymes of said library being classified by said enzyme characteristics, each of said at least three different selected enzymes each having at least two common enzyme characteristics.
placing in a package at least three different selected enzymes, said enzymes being selected from a recombinant enzyme library having a plurality of differentenzymes having different enzyme characteristics, said enzymes of said library being classified by said enzyme characteristics, each of said at least three different selected enzymes each having at least two common enzyme characteristics.
5. A process for identifying clones of a recombinant library which express a protein with a desired activity, comprising:
screening in the liquid phase a library of expression clones randomly produced from DNA of at least one microorganism, said screening being effected on expression products of said clones to thereby identify clones which express a protein with a desired activity.
screening in the liquid phase a library of expression clones randomly produced from DNA of at least one microorganism, said screening being effected on expression products of said clones to thereby identify clones which express a protein with a desired activity.
6. The process of claim 5 wherein the DNA from the library of expression clones is produced is gene cluster DNA.
7. The process of claim 5 wherein said protein is an enzyme.
8. A process of screening clones having DNA from an uncultivated microorganism for a specified protein activity, which process comprises:
screening for a specified protein activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein activity.
screening for a specified protein activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein activity.
9. The process of claim 8 wherein the recovered DNA is amplified.
10. The process of claim 8 wherein the recovered DNA is ligated into a vector.
11. The process of claim 10 wherein the vector into which the recovered DNA is ligated comprises at least one DNA sequence capable of regulating production of a detectable enzyme activity from said recovered DNA.
12. The process of claim 8 wherein the vector into which the recovered DNA has been ligated is used to transform a host.
13. The process of claim 8 wherein the protein is an enzyme.
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/503,606 | 1995-07-18 | ||
US08/503,606 US6004788A (en) | 1995-07-18 | 1995-07-18 | Enzyme kits and libraries |
US56899495A | 1995-12-07 | 1995-12-07 | |
US08/568,994 | 1995-12-07 | ||
US08/657,409 US5958672A (en) | 1995-07-18 | 1996-06-03 | Protein activity screening of clones having DNA from uncultivated microorganisms |
US08/657,904 | 1996-06-03 | ||
PCT/US1996/011854 WO1997004077A1 (en) | 1995-07-18 | 1996-07-17 | Screening methods for enzymes and enzyme kits |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2227342A1 true CA2227342A1 (en) | 1997-02-06 |
CA2227342C CA2227342C (en) | 2011-03-29 |
Family
ID=27054566
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2227342A Expired - Lifetime CA2227342C (en) | 1995-07-18 | 1996-07-17 | Screening methods for enzymes and enzyme kits |
Country Status (10)
Country | Link |
---|---|
US (8) | US5958672A (en) |
EP (2) | EP1696025B1 (en) |
JP (1) | JP2002514894A (en) |
AT (2) | ATE465246T1 (en) |
AU (1) | AU6547796A (en) |
CA (1) | CA2227342C (en) |
DE (2) | DE69638168D1 (en) |
DK (2) | DK0839185T3 (en) |
ES (1) | ES2277346T3 (en) |
WO (1) | WO1997004077A1 (en) |
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EP0839185A4 (en) | 2002-10-30 |
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