US4585739A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Plasmid for foreign gene expression in B. subtilis
Assignee: University of Maryland, College Park
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Filing: 1983-03-07
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Granted: 1986-04-29
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Expires: 2003-03-07
Abstract
Abstract The invention provides a mutant plasmid useful for introducing into B. subtilis foreign DNA, the nucleic acid sequence of which codes for production of a desired product. The plasmid is constructed from (1) a segment of B. pumllus DNA containing a chloramphenicol acetyltransferase gene plus an EcoRI* promoter fragment cloned from phage SPO2 DNA and a (2) portion of a B. subtilis plasmid, pUB110. The plasmid gives enhanced expression of genes coding for certain products, such as dihydrofolate reductase.
Inventors
- 👤 Paul S. Lovett
- 👤 Ronald G. Schoner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 20:51
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/74; C12N15/75; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/555; C07K14/565
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
105 words
A plasmid useful for introducing into B. subtilis foreign DNA, the nucleic acid sequence of which codes for production of a desired product, said plasmid being characterized as follows: (a) it is comprised of (1) a double-stranded DNA segment of B. pumilus strain NCIB8600 containing a chloramphenicol acetyltransferase gene plus an EcoRI* promoter fragment cloned from phage SPO2 DNA and (2) a portion of a B. subtilis plasmid pUB110; and (b) it has a mutation in its EcoRI to PstI fragment downstream from the promoter fragment whereby the nucleotide sequence for this EcoRI to PstI fragment is that shown in FIG. 1 of the drawing.
Claim 2
Dependent Claim
105 words
A plasmid useful for introducing into B. subtilis foreign DNA, the nucleic acid sequence of which codes for production of a desired product, said plasmid being characterized as follows: (a) it is comprised of (1) a double-stranded DNA segment of B. pumilus strain NCIB8600 containing a chloramphenicol acetyltransferase gene plus an EcoRI* promoter fragment cloned from phage SPO2 DNA and (2) a portion of a B. subtilis plasmid pUB110; and (b) it has a mutation in its EcoRI to PstI fragment downstream from the promoter fragment whereby the nucleotide sequence for this EcoRI to PstI fragment is that shown in FIG. 1 of the drawing.
Claim 3
Dependent Claim
17 words
A plasmid according to claim 1 wherein the foreign DNA codes for the production of human interferon.
Claim 4
Dependent Claim
12 words
A plasmid according to claim 2 wherein the human interferon is β-interferon.
Claim 5
Dependent Claim
30 words
A plasmid according to claim 1 wherein the B. pumilus segment is about 0.8 Md in molecular weight and the promoter fragment has a molecular weight of about 0.2 Md.
Claim 6
Dependent Claim
17 words
A plasmid according to claim 4 wherein the plasmid has a molecular weight of about 3.3 Md.
Claim 7
Dependent Claim
18 words
A plasmid according to claim 4 wherein the foreign DNA is a mouse gene coding for dihydrofolate reductase.
Claim 8
Dependent Claim
24 words
Plasmid pSA3-12 characterized as shown by endonuclease restriction map of FIG. 3 of the drawing and having a molecular weight of about 4.56 Md.
Claim 9
Dependent Claim
24 words
Plasmid pSA3-12 characterized as shown by endonuclease restriction map of FIG. 3 of the drawing and having a molecular weight of about 4.56 Md.
Claim 10
Dependent Claim
24 words
Plasmid pSA-XΔB characterized as shown by endonuclease restriction map of FIG. 4 of the drawing and having a molecular weight of about 4.56 Md.
Claim 11
Dependent Claim
24 words
Plasmid pSA-XΔB characterized as shown by endonuclease restriction map of FIG. 4 of the drawing and having a molecular weight of about 4.56 Md.
Claim 12
Dependent Claim
12 words
B. subtilis strain BGSCIS53 (pPL623), having the deposit accession number ATCC 39294.
Claim 13
Dependent Claim
12 words
B. subtilis strain BGSCIS53 (pPL623), having the deposit accession number ATCC 39294.
Claim 14
Dependent Claim
12 words
B. subtilis strain BGSCIS53 (pSA3-12), having the deposit accession number ATCC 39292.
Claim 15
Dependent Claim
12 words
B. subtilis strain BGSCIS53 (pSA3-12), having the deposit accession number ATCC 39292.
Claim 16
Dependent Claim
13 words
B. subtilis strain BGSCIS53 (pSA XΔB), having the deposit accession number ATCC 39293.
Claim 17
Dependent Claim
13 words
B. subtilis strain BGSCIS53 (pSA XΔB), having the deposit accession number ATCC 39293.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-04-28 | 1989-10-28 | 1990-04-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-04-28 | 1993-10-28 | 1994-04-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-04-28 | 1997-10-28 | 1998-04-29 | $7700.00 | Window Closed / Lapsed |