Claims
- 1. A method for enriching for target DNA sequences containing at least a partial coding region for at least one specified activity in a DNA sample comprising:
a) co-encapsulating in a micro-environment a mixture of target DNA obtained from more than one organism with a mixture of DNA probes comprising a detectable marker and at least a portion of a DNA sequence encoding at least one enzyme having a specified enzyme activity; b) incubating the co-encapsulated mixture under such conditions and for such time as to allow hybridization of complementary sequences; and c) screening for the specified activity.
- 2. The method of claim 1, further comprising transforming host cells with recovered target DNA to produce an expression library of a plurality of clones.
- 3. The method of claim 1, wherein the organisms are microorganisms.
- 4. The method of claim 3, wherein the microorganisms are uncultured microorganisms.
- 5. The method of claim 1, further comprising screening the expression library for the specified enzyme activity.
- 6. 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 c) transforming a host cell with the double stranded DNA of c).
- 7. A method of FACS screening for an agent that modulates the activity of a target cell component, wherein the target cell component and a selectable marker are expressed by a eukaryotic cell, the method comprising co-encapsulating the agent in a microenvironment with the recombinant cell expressing the target cell component and detectable marker and detecting the effect of the agent on the activity of the cell component.
- 8. The method of claim 1, wherein said target DNA is gene cluster DNA.
- 9. The method of claim 4, wherein the uncultured microorganisms are derived from an environmental sample.
- 10. The method of claim 4, 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.
- 11. The method of claim 2, wherein the clones comprise a construct selected from the group consisting of phage, plasmids, phagemids, cosmids, fosmids, viral vectors, and artificial chromosomes.
- 12. The method of claim 1, wherein the target DNA comprises one or more operons, or portions thereof, of the DNA population.
- 13. The method of claim 12, wherein the operon or portions thereof encodes a complete or partial metabolic pathway.
- 14. The method of claim 4, wherein the uncultured microorganisms comprise extremophiles.
- 15. The method of claim 14, wherein the extremophiles are selected from the group consisting of thermophiles, hyperthermophiles, psychrophiles, barophiles, and psychrotrophs.
- 16. The method of claim 6, wherein the host cell is selected from the group consisting of a bacterium, fungus, plant cell, insect cell and animal cell.
- 17. The method of claim 1, wherein the target DNA encodes a protein.
- 18. The method of claim 17, wherein the protein is an enzyme.
- 19. The method of claim 18, wherein the enzyme is selected from the group consisting of oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases.
- 20. The method of claim 1, wherein the micro-environment is a liposome, gel microdrop, bead, agarose, cell, ghost red blood cell or ghost macrophage.
- 21. The method of claim 20, wherein the liposomes are prepared from one or more phospholipids, glycolipids, steroids, alkyl phosphates or fatty acid esters.
- 22. The method of claim 21, wherein the phospholipids are selected from the group consisting of lecithin, sphingomyelin and dipalmitoyl.
- 23. The method of claim 20, wherein the steroids are selected from the group consisting of cholesterol, cholestanol and lanosterol.
- 24. The method of claim 1, wherein the detectable marker is a fluorescent dye, a visible dye, a bioluminescent material, a chemiluminescent material, a radioactive material, or an enzymatic substrate.
- 25. The method of claim 24, wherein the bioluminescent material is green fluorescent protein (GFP) or red fluorescent protein (RFP).
- 26. The method of claim 25, wherein detection of the fluorescent dye or a visible dye is carried out by fluorometric or spectrophotometric measurement.
Parent Case Info
[0001] This application is a divisional of U.S. patent application Ser. No. 09/636,778, filed Aug. 11, 2000, which is a continuation of U.S. patent application Ser. No. 09/098,206, filed Jun. 16, 1998, continuation-in-part of U.S. application Ser. No. 08/876,276, filed Jun. 16, 1997, now abandoned.
Divisions (1)
|
Number |
Date |
Country |
Parent |
09636778 |
Aug 2000 |
US |
Child |
09848185 |
May 2001 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09098206 |
Jun 1998 |
US |
Child |
09636778 |
Aug 2000 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
08876276 |
Jun 1997 |
US |
Child |
09098206 |
Jun 1998 |
US |