US5912119A
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Process for the in vivo recombination of DNA sequences having mismatched bases
Assignee: Mixis France SA
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Filing: 1994-04-25
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Granted: 1999-06-15
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Expires: 2014-04-25
Abstract
Abstract The present invention relates to a process of recombination in vivo of partially homologous DNA sequences having up to 30% of base mismatches. According to its essential characteristic, said sequences are placed together in cells or an organism of which the enzymatic mismatch repair system is defective or has been transitorily inactivated by saturation for the time to obtain recombination between said DNA sequences or in using mutants which increase the intergeneric recombination.
Inventors
- 👤 Miroslav Radman
- 👤 Christiane Rayssiguier
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 21:20
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/102; C12N15/1027
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
325 words
A process for in vivo intergeneric and/or interspecific recombination, comprising: combining in a cell a double-stranded DNA from a first species or genus with a double stranded DNA from a second species or genus, said first and second DNAs having sequences which are partially homologous and have mismatches able to activate the enzymatic mismatch repair system of the cell when said system is functional, wherein the enzymatic mismatch repair system is defective or has been inactivated transitorily to enable stable recombination between said DNA sequences, wherein said process is selected from the group of processes consisting of: (i) fusing cells of an organism of a first species or a first genus with cells of an organism of a second species or a second genus, said cells of said organism of a second species or a second genus having a defective enzymatic mismatch repair system or having said system inactivated transitorily; (ii) crossing a unicellular organism of a first species or a first genus with a unicellular organism of a second species or a second genus, wherein at least one organism has a defective enzymatic mismatch repair system or has said system inactivated transitorily; (iii) conjugating or transducting a recipient bacterium of a first species or a first genus and a donor bacterium of a second species or a second genus, said donor bacterium having at least one DNA sequence to be transferred to said recipient bacterium, wherein at least one of said donor bacterium and said recipient bacterium includes a defective enzymatic mismatch repair system or said system is inactivated transitorily; (iv) inactivating said enzymatic mismatch repair system by at least one mutation of at least one of the mutS and mutL genes; and (v) placing into a unicellular organism defective or inactivated in the enzymatic mismatch repair system two DNA sequences, said two DNA sequences being partially homologous genes derived from two different organisms, and selecting the desired hybrid sequence, gene, or product thereof.
Claim 2
Dependent Claim
325 words
A process for in vivo intergeneric and/or interspecific recombination, comprising: combining in a cell a double-stranded DNA from a first species or genus with a double stranded DNA from a second species or genus, said first and second DNAs having sequences which are partially homologous and have mismatches able to activate the enzymatic mismatch repair system of the cell when said system is functional, wherein the enzymatic mismatch repair system is defective or has been inactivated transitorily to enable stable recombination between said DNA sequences, wherein said process is selected from the group of processes consisting of: (i) fusing cells of an organism of a first species or a first genus with cells of an organism of a second species or a second genus, said cells of said organism of a second species or a second genus having a defective enzymatic mismatch repair system or having said system inactivated transitorily; (ii) crossing a unicellular organism of a first species or a first genus with a unicellular organism of a second species or a second genus, wherein at least one organism has a defective enzymatic mismatch repair system or has said system inactivated transitorily; (iii) conjugating or transducting a recipient bacterium of a first species or a first genus and a donor bacterium of a second species or a second genus, said donor bacterium having at least one DNA sequence to be transferred to said recipient bacterium, wherein at least one of said donor bacterium and said recipient bacterium includes a defective enzymatic mismatch repair system or said system is inactivated transitorily; (iv) inactivating said enzymatic mismatch repair system by at least one mutation of at least one of the mutS and mutL genes; and (v) placing into a unicellular organism defective or inactivated in the enzymatic mismatch repair system two DNA sequences, said two DNA sequences being partially homologous genes derived from two different organisms, and selecting the desired hybrid sequence, gene, or product thereof.
Claim 3
Dependent Claim
57 words
The process of claim 1 wherein there is produced a hybrid gene and its encoded protein, by placing into a unicellular organism defective or inactivated in the enzymatic mismatch repair system two DNA sequences, said two DNA sequences being partially homologous genes derived from two different organisms, and selecting the desired hybrid gene or its coded protein.
Claim 4
Dependent Claim
36 words
The process of part (v) of claim 1, wherein each DNA sequence is contained within a separate plasmid, wherein each plasmid is introduced into a bacterium which includes a defective or inactivated mismatched DNA repair system.
Claim 5
Dependent Claim
66 words
The process of claim 1 comprising combining (i) DNA of cells of an organism of a first species or a first genus with (ii) chromosomal DNA of cells of an organism of a second species or a second genus, said cells of an organism of a second species or a second genus being defective in the enzymatic mismatch repair system or having said system inactivated transitorily.
Claim 6
Dependent Claim
64 words
The process of claim 1 wherein said process comprises a fusion of cells of an organism of a first species or a first genus and of cells of an organism of a second species or a second genus, said cells of said organism of a second species or a second genus having a defective enzymatic mismatch repair system or having said system inactivated transitorily.
Claim 7
Dependent Claim
53 words
The process of claim 1 wherein said process comprises a cross between a unicellular organism of a first species or a first genus and a unicellular organism of a second species or a second genus, wherein at least one organism has a defective enzymatic mismatch repair system or having said system inactivated transitorily.
Claim 8
Dependent Claim
79 words
The process of claim 1 wherein said process comprises a conjugation or a transduction between a recipient bacterium of a first species or a first genus and a donor bacterium of a second species or a second genus, said donor bacterium having at least one DNA sequence to be transferred to said recipient bacterium, wherein at least one of said donor bacterium and said recipient bacterium includes a defective enzymatic mismatch repair system or said system is inactivated transitorily.
Claim 9
Dependent Claim
22 words
The process of claim 1 wherein said enzymatic mismatch repair system is inactivated transitorily by saturation of said system with non-homologous DNA.
Claim 10
Dependent Claim
28 words
The process of claim 1 wherein said enzymatic mismatch repair system is inactivated by at least one mutation of at least one of the mutS and mutL genes.
Claim 11
Dependent Claim
17 words
The process of claim 1 wherein said partially homologous DNA sequences have up to 30% of mismatches.
Claim 12
Dependent Claim
25 words
The process of claim 1, wherein in (i), said cells are selected from the group of cells consisting of bacteria, yeast, plant, and animal cells.
Claim 13
Dependent Claim
321 words
A process for intergeneric and/or interspecific recombination, comprising: combining in a cell a DNA from a first species or genus with a DNA from a second species or genus, wherein said first and second DNAs have sequences which are partially homologous and have mismatches able to activate the enzymatic mismatch repair system of the cell when said system is functional, wherein the enzymatic mismatch repair system is defective or has been inactivated transitorily to enable stable recombination between said DNA sequences, wherein said process is selected from the group of processes consisting of: (i) fusing cells of an organism of a first species or a first genus with cells of an organism of a second species or a second genus, said cells of said organism of a second species or a second genus having a defective enzymatic mismatch repair system or having said system inactivated transitorily; (ii) crossing a unicellular organism of a first species or a first genus with a unicellular organism of a second species or a second genus, wherein at least one organism has a defective enzymatic mismatch repair system or has said system inactivated transitorily; (iii) conjugating or transducing a recipient bacterium of a first species or a first genus and a donor bacterium of a second species or a second genus, said donor bacterium having at least one DNA sequence to be transferred to said recipient bacterium, wherein at least one of said donor bacterium and said recipient bacterium includes a defective enzymatic mismatch repair system or said system is inactivated transitorily; (iv) inactivating said enzymatic mismatch repair system by at least one mutation of at least one of the mutS and mutL genes; and (v) placing into a unicellular organism defective or inactivated in the enzymatic mismatch repair system two DNA sequences, said two DNA sequences being partially homologous genes derived from two different organisms, and selecting the desired hybrid sequence, gene, or product thereof.
Claim 14
Dependent Claim
321 words
A process for intergeneric and/or interspecific recombination, comprising: combining in a cell a DNA from a first species or genus with a DNA from a second species or genus, wherein said first and second DNAs have sequences which are partially homologous and have mismatches able to activate the enzymatic mismatch repair system of the cell when said system is functional, wherein the enzymatic mismatch repair system is defective or has been inactivated transitorily to enable stable recombination between said DNA sequences, wherein said process is selected from the group of processes consisting of: (i) fusing cells of an organism of a first species or a first genus with cells of an organism of a second species or a second genus, said cells of said organism of a second species or a second genus having a defective enzymatic mismatch repair system or having said system inactivated transitorily; (ii) crossing a unicellular organism of a first species or a first genus with a unicellular organism of a second species or a second genus, wherein at least one organism has a defective enzymatic mismatch repair system or has said system inactivated transitorily; (iii) conjugating or transducing a recipient bacterium of a first species or a first genus and a donor bacterium of a second species or a second genus, said donor bacterium having at least one DNA sequence to be transferred to said recipient bacterium, wherein at least one of said donor bacterium and said recipient bacterium includes a defective enzymatic mismatch repair system or said system is inactivated transitorily; (iv) inactivating said enzymatic mismatch repair system by at least one mutation of at least one of the mutS and mutL genes; and (v) placing into a unicellular organism defective or inactivated in the enzymatic mismatch repair system two DNA sequences, said two DNA sequences being partially homologous genes derived from two different organisms, and selecting the desired hybrid sequence, gene, or product thereof.
Claim 15
Dependent Claim
25 words
The process of claim 12, wherein in (i), said cells are selected from the group of cells consisting of bacteria, yeast, plant, and animal cells.
References Cited (Backward Citations)
No citation records indexed.
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