US4839293A
Expired / Lapsed
US
⏱️ 0.0 years remaining
DNA encoding streptavidin, streptavidin produced therefrom, fused polypeptides which include amino acid sequences present in streptavidin and uses thereof
Assignee: Columbia University
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Filing: 1986-02-24
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Granted: 1989-06-13
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Expires: 2006-02-24
Abstract
Abstract DNA which encodes the polypeptide streptavidin has been isolated as a fragment 2 kb in length derived from a restriction endonuclease digestion of the chromosomal DNA of Streptomyces avidinii. The nucleic acid sequence of the gene and the amino acid sequence of the polypeptide have been determined. A fused gene has been prepared which comprises the streptavidin gene fused to a gene encoding the human LDL receptor. Expression of the gene fusion results in a fused streptavidin-human LDL receptor polypeptide. Methods are provided for using the fused gene to produce labeled, chemically modified proteins in vivo and to isolate a protein knowing only the nucleotide sequence of the gene encoding the protein.
Inventors
- 👤 Charles R. Cantor
- 👤 Richard Axel
- 👤 Carlos Argarana
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 18:20
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/195; C07K14/36; C07K14/435; C07K14/705; C07K2319/00; C07K2319/20; C07K2319/22; C07K2319/33; Y; Y10; Y10S; Y10S930/00; Y10S930/01; Y10S930/30
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
18 words
An isolated DNA molecule encoding streptavidin and comprising the nucleic acid coding sequence set forth in FIG. 3.
Claim 2
Dependent Claim
18 words
An isolated DNA molecule encoding streptavidin and comprising the nucleic acid coding sequence set forth in FIG. 3.
Claim 3
Dependent Claim
16 words
A recombinant cloning vehicle which comprises a cloning vehicle DNA and the DNA of claim 1.
Claim 4
Dependent Claim
39 words
The cloning vehicle of claim 2, comprising the entire region coding for the polypeptide streptavidin, a region encoding a signal peptide and the flanking region DNA which occurs naturally at the 3' and 5' ends of the coding region.
Claim 5
Dependent Claim
7 words
A plasmid cloning vehicle of claim 2.
Claim 6
Dependent Claim
7 words
A phage cloning vehicle of claim 2.
Claim 7
Dependent Claim
16 words
The phage cloning vehicle of claim 5, wherein the cloning vehicle is derived from phage M13.
Claim 8
Dependent Claim
17 words
The plasmid cloning vehicle of claim 4, wherein the cloning vehicle is derived from the pUC plasmid.
Claim 9
Dependent Claim
14 words
A genetically engineered bacterial host cell which comprises the cloning vehicle of claim 2.
Claim 10
Dependent Claim
8 words
An E. coli host cell of claim 8.
Claim 11
Dependent Claim
16 words
An E. coli host cell of claim 9 designated JM83 and having ATCC Accession No. 53307.
Claim 12
Dependent Claim
12 words
A mammalian host cell which comprises the cloning vehicle of claim 2.
Claim 13
Dependent Claim
16 words
A mammalian host cell of claim 11 wherein the mammalian cell is a NIH 3T3 cell.
Claim 14
Dependent Claim
69 words
A fused gene which comprises a first DNA molecule encoding a protein fused to said streptavidin encoding DNA molecule of claim 1, wherein the streptavidin has a multiplicity of binding sites for biotin or a biotin derivative, and wherein the fused gene is capable of expressing a fused protein in vivo when said fused gene is inserted into a suitable expression vector and introduced into a suitable host cell.
Claim 15
Dependent Claim
20 words
The fused gene of claim 13 wherein the first DNA molecule is at the 5' end of the fused gene.
Claim 16
Dependent Claim
21 words
The fused gene of claim 13 wherein the DNA molecule encoding streptavidin is at the 5' end of the fused gene.
Claim 17
Dependent Claim
20 words
The fused gene of claim 13, wherein the first DNA molecule is the gene encoding the human LDL receptor protein.
Claim 18
Dependent Claim
53 words
The fused gene of claim 16, which is capable of expressing protein that consists of streptavidin at the N-terminal region of the fused protein and the LDL receptor protein at the C-terminal region of the fused protein when the fused gene is inserted into an expression vector and introduced into a host cell.
Claim 19
Dependent Claim
39 words
An expression vector capable of expressing the fused gene of claim 13, when introduced into a host cell, which comprises carrier DNA and said fused gene of claim 13 wherein said carrier DNA comprises a promoter and operator region.
Claim 20
Dependent Claim
7 words
A mammalian expression vector of claim 18.
Claim 21
Dependent Claim
12 words
A mammalian host cell which comprises the expression vector of claim 19.
Claim 22
Dependent Claim
8 words
An NIH 3T3 host cell of claim 20.
Claim 23
Dependent Claim
30 words
An expression vector capable of expressing the fused gene of claim 16, when introduced into a host cell, which comprises carrier DNA and the fused DNA fragments of claim 16.
Claim 24
Dependent Claim
7 words
A mammalian expression vector of claim 22.
Claim 25
Dependent Claim
12 words
A mammalian host cell which comprises the expression vector of claim 23.
Claim 26
Dependent Claim
8 words
An NIH 3T3 host cell of claim 24.
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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) | 1992-06-12 | 1992-12-12 | 1993-06-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-06-12 | 1996-12-12 | 1997-06-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-06-12 | 2000-12-12 | 2001-06-13 | $7700.00 | Window Closed / Lapsed |