US4649109A
Expired / Lapsed
US
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Methods for isolating mutant microorganisms from parental populations
Assignee: Brandeis University
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Filing: 1984-02-16
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Granted: 1987-03-10
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Expires: 2004-02-16
Abstract
Abstract A method for isolating a mutant microorganism is described. The method comprises the steps of: (a) separately microencapsulating in a semi-permeable membrane each or a small number of microorganisms from a microorganism population containing said mutant; (b) growing said microencapsulated microorganisms including treating to induce a detectable difference between microcapsules containing mutant microorganisms and those containing non-mutant microorganisms; and (c) separating said microcapsules containing mutant microorganisms from those containing non-mutant microorganisms based on said difference.
Inventors
- 👤 Daniel Perlman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 12:05
CPC Classifications
C; C12; C12N; C12N7/00; C12N11/00; C12N11/02; C12N11/04; C12N15/00; C12Q; C12Q1/00; C12Q1/02; C12Q1/24; C12N2710/00; C12N2710/00011; C12N2710/00051
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
82 words
A method for isolating a mutant microorganism, which method comprises the steps of: a. separately microencapsulating in a semi-permeable membrane each or a small number of microorganisms from a microorganism population containing said mutant; b. growing said microencapsulated microorganisms and treating them to induce a difference in cell number between microcapsules containing mutant microorganisms and those containing non-mutant microorganisms; and c. separating said microcapsules containing mutant micoorganisms from those containing non-mutant microorganisms by a method based on said difference in cell numbers.
Claim 2
Independent Claim
82 words
A method for isolating a mutant microorganism, which method comprises the steps of: a. separately microencapsulating in a semi-permeable membrane each or a small number of microorganisms from a microorganism population containing said mutant; b. growing said microencapsulated microorganisms and treating them to induce a difference in cell number between microcapsules containing mutant microorganisms and those containing non-mutant microorganisms; and c. separating said microcapsules containing mutant micoorganisms from those containing non-mutant microorganisms by a method based on said difference in cell numbers.
Claim 3
Dependent Claim
19 words
The method of claim 1, wherein said mutant microorganism is a naturally occuring mutant of a wild type microorganism.
Claim 4
Dependent Claim
15 words
The method of claim 1, wherein the mutation in the mutant microorgahism is artificially induced.
Claim 5
Dependent Claim
22 words
The method of claim 3, wherein said artificially-induced mutant is the result of genetic engineering by a biological, biochemical or biophysical process.
Claim 6
Dependent Claim
67 words
The method of claim 1, wherein said microcapsules containing said single microorganisms are formed by: a. forming a dilute suspension of said microorganisms in a liquid diluent capable of forming a gel upon said dilute suspension having a dilution selected so that there is a high probability that each microcapsule produced from said suspension contains one microorganism; b. converting said suspension into gel droplets, forming said microcapsules.
Claim 7
Dependent Claim
34 words
The method of claim 1, wherein said treatment comprises growing said microencapsulated microorganisms under conditions restrictive to said mutant whereby said microcapsules containing said mutants have fewer microorganisms per microcapsule than microcapsules containing non-mutants.
Claim 8
Dependent Claim
50 words
In the method of claim 6, prior to said separation step, incubating said microcapsules in a high-density medium of appropriate diffusion rate, thereby causing the mutants in the microcapsules which have fewer microorganisms to have a higher density than the non-mutants in the microcapsules which have larger numbers of microorganisms.
Claim 9
Dependent Claim
37 words
The method of claim 1 wherein in said growing step (b) the microencasulated mutants have a capacity to accumulate a particular material resulting in an increase in mass and number greater than that of said microencapsulated non-mutants.
Claim 10
Dependent Claim
45 words
The method of claim 1, wherein said separation is based on the difference in mass between said microcapsules containing mutant and non-mutant microorganisms and comprises (a) equilibrium density centrifugation of a suspension containing said microcapsules or (b) velocity sedimentation of a suspension containing said microcapsules.
Claim 11
Dependent Claim
11 words
The method of claim 1, wherein said microorganisms comprise eucaryotic cells.
Claim 12
Dependent Claim
11 words
The method of claim 1, wherein said microorganisms comprise procaryotic cells.
Claim 13
Dependent Claim
10 words
The method of claim 1, wherein said microorganisms comprise viruses.
Claim 14
Dependent Claim
11 words
The method of claim 1, wherein said microorganisms comprise hybridoma cells.
Claim 15
Dependent Claim
22 words
The method of claim 1, wherein mutant microorganisms are selected from chemically complex and non-sterile agricultural, industrial or other commercial process environments.
Claim 16
Dependent Claim
16 words
The method of claim 1, wherein said microencapsulated microorganisms are grown in a laboratory culturing medium.
Claim 17
Dependent Claim
32 words
The method of claim 1, wherein said microencapsulated microorganisms are grown in an agricultural or industrial process medium wherein one or more species of said microorganism are found in the native environment.
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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) | 1990-03-09 | 1990-09-08 | 1991-03-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-03-09 | 1994-09-08 | 1995-03-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-03-09 | 1998-09-08 | 1999-03-10 | $7700.00 | Window Closed / Lapsed |