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EP0072925A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

T4 DNA fragment as a stabilizer for proteins expressed by cloned DNA

Assignee: Rutgers Research and Educational Foundation
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Filing: 1982-07-21
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Granted: —
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Expires: 2002-07-21
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Abstract

Abstract A DNA fragment isolated from the DNA of T-even bacteriophage which suppresses the degradation of abnormal proteins in bacterial cells is disclosed. The DNA fragment can be isolated by treating the DNA of T-even bacteriophage with a restriction enzyme to cleave the DNA into fragments, at least one of said fragments consisting essentially of a sequence of nucleotide bases which code for protein that functions to suppress degradation of abnormal proteins in bacterial cells, inserting the DNA fragments into cloning vehicles, such as plasmids, to produce hybrid cloning vehicles, transforming bacterial cells by introducing the hybrid cloning vehicles therein, and selecting the transformants which exhibit suppressed protein degradation mechanisms. The DNA fragment can then be isolated from the selected transformants. The stabilizing DNA fragment can be introduced into bacterial host cells to increase the yield of foreign proteins expressed by cloned DNA.

Inventors

  • 👤 Lee D. Simon
  • 👤 Rose B. Fay

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-10-25 23:56
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; C07K14/435; C07K14/52; C07K14/555; C07K14/565; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/70; C12N2795/00; C12N2795/00011; C12N2795/10011; C12N2795/10111; C12N2795/10122

Claims Analysis

Total Claims: 20
Claim 1
Independent Claim 36 words
A DNA fragment isolated from the DNA of a T-even bacteriophage, said D NA fragment consisting essentially of a sequence of nucleotide bases which code for protein that suppresses degradation of abnormal polypeptides in bacterial cells.
Claim 2
Dependent Claim 36 words
A DNA fragment isolated from the DNA of a T-even bacteriophage, said D NA fragment consisting essentially of a sequence of nucleotide bases which code for protein that suppresses degradation of abnormal polypeptides in bacterial cells.
Claim 3
Dependent Claim 14 words
The DNA fragment of Claim 1 wherein said fragment is isolated from T4 bacteriophage.
Claim 4
Dependent Claim 14 words
The DNA fragment of Claim 1 wherein said T-even DNA is cytosine-containing and nonglucosylated.
Claim 5
Dependent Claim 15 words
A hybrid cloning vehicle comprising a cloning vehicle and the DNA fragment of Claim 1.
Claim 6
Dependent Claim 14 words
The hybrid cloning vehicle of Claim 4 wherein said cloning vehicle is a plasmid.
Claim 7
Dependent Claim 25 words
The hybrid cloning vehicle of Claim 4 wherein said cloning vehicle further comprises a sequence of nucleotide bases coding for a eukaryotic preferably human protein.
Claim 8
Dependent Claim 14 words
The hybrid cloning vehicle of Claim 6 wherein said human protein is human interferon.
Claim 9
Dependent Claim 12 words
A bacterial cell containing the hybrid cloning vehicle of Claim 6. '
Claim 10
Dependent Claim 33 words
A bacterial cell containing the hybrid cloning vehicle of Claim 4 and further containing a second cloning vehicle, said second cloning vehicle containing a sequence of nucleotide bases coding for a eukaryotic protein.
Claim 11
Dependent Claim 47 words
A method for suppressing the degradation of abnormal proteins in a bacterial cell, comprising producing the DNA fragment of Claim 1, inserting said DNA fragment into a cloning vehicle to produce a hybrid cloning vehicle, and transforming a bacterial cell by introducing said hybrid cloning vehicle therein.
Claim 12
Independent Claim 92 words
A method for producing a DNA fragment consisting essentially of a sequence of nucleotide bases coding for protein that suppresses degradation of abnormal proteins in bacterial cells, comprising treating the DNA of a T-even bacteriophage with a restriction enzyme to cleave the DNA into fragments, at least one of said fragments coding for protein that suppresses degradation in bacterial cells, inserting said DNA fragments into cloning vehicles to produce hybrid cloning vehicles, transforming bacterial cells by introducing said hybrid cloning vehicles therein, and selecting the transformants exhibiting suppressed degradation of abnormal proteins..
Claim 13
Independent Claim 92 words
A method for producing a DNA fragment consisting essentially of a sequence of nucleotide bases coding for protein that suppresses degradation of abnormal proteins in bacterial cells, comprising treating the DNA of a T-even bacteriophage with a restriction enzyme to cleave the DNA into fragments, at least one of said fragments coding for protein that suppresses degradation in bacterial cells, inserting said DNA fragments into cloning vehicles to produce hybrid cloning vehicles, transforming bacterial cells by introducing said hybrid cloning vehicles therein, and selecting the transformants exhibiting suppressed degradation of abnormal proteins..
Claim 14
Dependent Claim 26 words
The method of Claim 11 further comprising removing said hybrid cloning vehicle from a selected transformant and excising said DNA fragment from said hybrid cloning vehicle.
Claim 15
Dependent Claim 23 words
A nucleic acid consisting essentially of a sequence of nucleotide bases coding for protein that suppresses degradation of abnormal proteins in bacterial cells.
Claim 16
Dependent Claim 23 words
A nucleic acid consisting essentially of a sequence of nucleotide bases coding for protein that suppresses degradation of abnormal proteins in bacterial cells.
Claim 17
Dependent Claim 8 words
A DNA fragment in accordance with Claim 13.
Claim 18
Dependent Claim 15 words
A hybrid cloning vehicle comprising a cloning vehicle and the DNA fragment of Claim 14.
Claim 19
Dependent Claim 11 words
A bacterial cell containing the hybrid cloning vehicle of Claim 15.
Claim 20
Dependent Claim 47 words
A method for suppressing the degradation of abnormal proteins in a bacterial cell comprising producing the DNA fragment of Claim 14, inserting said DNA fragment into a cloning vehicle to produce a hybrid cloning vehicle, and transforming a bacterial cell by introducing said hybrid cloning vehicle therein.

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