EP0076037A2
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EP
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Amplified expression of DNA sequences
Assignee: Biogen Inc
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Filing: 1982-08-26
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Granted: —
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Expires: 2002-08-26
Abstract
Abstract A process for the amplified expression of DNA sequences and for the improved production of amino acids, polypeptides and proteins coded for by those DNA sequences. The process comprises the steps of increasing the copy number per cell of a multicopy plasmid-carried DNA sequence coding for a desired product; combining culture media with a culture of those increased copy number cells; culturing the resulting culture under conditions appropriate for expression of the plasmid-carried DNA sequence; and harvesting the resulting culture before substantial exponential growth has occurred. The process affords high yields of those products, low contamination of those products and ease in fermentation culture monitoring, handling and purification.
Inventors
- 👤 Hans Kupper
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 22:14
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/005; C12; C12N; C12N1/00; C12N1/38; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/69; C12N2730/00; C12N2730/00011; C12N2730/10011; C12N2730/10111; C12N2730/10122
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
83 words
A process for amplifying the expression of multicopy plasmid-carried DNA sequences and improving the production of the products coded for by those DNA sequences comprising the steps of increasing the copy number per cell of a multicopy plasmid-carried DNA sequence coding for a desired product; combining culture media with a culture of said increased copy number cells; culturing the resultant culture under conditions appropriate for expression of the multicopy plasmid-carried DNA sequence; and harvesting the resulting culture before substantial exponential growth has occurred.
Claim 2
Dependent Claim
83 words
A process for amplifying the expression of multicopy plasmid-carried DNA sequences and improving the production of the products coded for by those DNA sequences comprising the steps of increasing the copy number per cell of a multicopy plasmid-carried DNA sequence coding for a desired product; combining culture media with a culture of said increased copy number cells; culturing the resultant culture under conditions appropriate for expression of the multicopy plasmid-carried DNA sequence; and harvesting the resulting culture before substantial exponential growth has occurred.
Claim 3
Dependent Claim
72 words
The process of claim 1 wherein the method of increasing the copy number per cell of a multicopy plasmid-carried DNA sequence coding for a desired product is selected from the group consisting of treating cells transformed with a multicopy plasmid-carried DNA sequence with chloramphenicol overnight and removing said chloramphenicol from said cells; and growing cells transformed with a multicopy plasmid-carried DNA sequence under conditions of amino acid starvation and combinations of them.
Claim 4
Dependent Claim
40 words
The process of claim 2 wherein the method of increasing the copy number per cell of a multicopy plasmid-carried DNA sequence coding for a desired product comprises growing an overnight culture of cells transformed with said multicopy plasmid-carried DNA sequence.
Claim 5
Dependent Claim
15 words
The process of claim 3 wherein said culture is grown for more than 18 h.
Claim 6
Dependent Claim
31 words
The process of any one of claims 1 to 4 wherein the culture media has a 0.05 to 10 times larger volume than the culture of said increased copy number cells.
Claim 7
Dependent Claim
25 words
The process of claim 5-wherein the culture media has a 0.05 to 0.2 times larger volume than the culture of said increased copy number cells.
Claim 8
Dependent Claim
26 words
The process of claim 5 wherein the culture media has a 3 to 5 times larger volume than the culture of said increased copy number cells.
Claim 9
Dependent Claim
30 words
The process of any one of claim 1 to 6 wherein the cell is selected from the group consisting of strains of E. coli, Bacillus, Pseudomonas, yeasts and other fungi.
Claim 10
Dependent Claim
12 words
The process of claim 8 wherein the cell is E. coli C600.
Claim 11
Dependent Claim
44 words
The process of any one of claims 1 to wherein said DNA sequence is selected from the group consisting of DNA sequences coding for active components of vaccines, pharmaceutical products, agricultural products, enzymes, hormones, polypeptides, proteins, amino acids, industrial chemicals, antibiotics and food stuffs.
Claim 12
Dependent Claim
77 words
The process of claim 10 wherein said DNA sequence is selected from the group consisting of DNA sequence coding for products displaying an immunological or biological activity of leukocyte interferon, products displaying an immunological or biological activity of fibroblast interferon, products displaying the antigenicity of FMD viral antigens, products displaying the biological activity of human or animal growth hormones, products displaying the biological activity of human insulin and products displaying the antigenicity of hepatitis B viral antigens.
References Cited (Backward Citations)
No citation records indexed.
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