Patent US4872133A successfully refreshed from Google Patents.
Patent EP0196864A2 successfully refreshed from Google Patents.
EP0196864A2
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EP
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Alkaline phosphatase-mediated processing and secretion of recombinant proteins, DNA sequences for use therein and cells transformed using such sequences
Assignee: Novartis Vaccines and Diagnostics Inc
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Filing: 1986-03-25
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Granted: —
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Expires: 2006-03-25
Abstract
Abstract @ Expression systems which are capable of secreting soluble, biologically active forms of proteins which are susceptible to processing in procaryotes under the influence of bacterial leader sequences is disclosed. Vectors successful in effecting this expression encode a fusion protein having an N-terminal sequence comprising the phoA (alkaline phosphatase) leader peptide and as a C-terminal sequence the desired protein. This fusion protein encoding sequence is placed under the control of a suitable bacterial promoter, preferably the alkaline phosphatase promoter. Terminator sequences may also be included in the vectors for efficient expression.
Inventors
- 👤 Shing Chang
- 👤 Leo Shun-Lee Lin
- 👤 Sheng-Yung Chang
- 👤 Alice Ming Wang
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 16:39
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/525; C07K14/575; C07K14/61; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N9/00; C12N9/14; C12N9/16; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/034; C07K2319/70; C07K2319/74; C07K2319/75
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
44 words
A recombinant DNA sequence useful in expression and secretion of a desired coding sequence in procaryotic cells which sequence encodes a fusion protein having as its N-terminal portion the leader sequence of alkaline phosphatase A (phoA) and as its C-terminal portion the desired peptide.
Claim 2
Dependent Claim
44 words
A recombinant DNA sequence useful in expression and secretion of a desired coding sequence in procaryotic cells which sequence encodes a fusion protein having as its N-terminal portion the leader sequence of alkaline phosphatase A (phoA) and as its C-terminal portion the desired peptide.
Claim 3
Dependent Claim
18 words
A DNA sequence as claimed in claim 1, wherein the desired protein is a protein susceptible to processing.
Claim 4
Dependent Claim
19 words
A DNA sequence as claimed in claim 1 or claim 2, wherein the desired protein is hGH or TNF..
Claim 5
Dependent Claim
13 words
A DNA sequence as claimed in claim 3 which is pSYC1053 or pAW721.
Claim 6
Dependent Claim
29 words
A DNA sequence as claimed in any one of claims 1 to 3 which further includes, in operable linkage with said DNA sequence, a promotor compatible with procaryotic hosts.
Claim 7
Dependent Claim
15 words
A DNA sequence as claimed in claim 5, wherein the promotor is the phoA promotor.
Claim 8
Dependent Claim
31 words
A DNA sequence as claimed in any one of claims 1 to 3, 5 or 6 which further includes, in operable linkage with said DNA sequence, a bacterially compatible terminator sequence.
Claim 9
Dependent Claim
27 words
A DNA sequence as claimed in claim 7, wherein the terminator is the phoA terminator or the terminator for the B . thuringiensis crystal protein gene (cry).
Claim 10
Dependent Claim
21 words
A DNA sequence as claimed in any one of claims 5 to 8 which is disposed on a bacterial cloning vector.
Claim 11
Dependent Claim
10 words
Recombinant host cells transformed with the vector of claim 9.
Claim 12
Dependent Claim
50 words
The use of the leader sequence of alkaline phosphatase A (phoA) as the N-terminal portion of a recombinant DNA sequence when linked to a desired coding sequence as the C-terminal portion of such recombinant DNA sequence in the expression of said desired coding sequence and secretion of the corresponding peptide.
Claim 13
Dependent Claim
50 words
The use of the leader sequence of alkaline phosphatase A (phoA) as the N-terminal portion of a recombinant DNA sequence when linked to a desired coding sequence as the C-terminal portion of such recombinant DNA sequence in the expression of said desired coding sequence and secretion of the corresponding peptide.
Claim 14
Dependent Claim
26 words
The use claimed in claim 11 when said recombinant DNA sequence is further defined by the specific feature of any one of claims 2 to 9.
Claim 15
Dependent Claim
20 words
A method for secreting a desired soluble biologically active protein which method comprises culturing cells as defined in claim 10.
Claim 16
Dependent Claim
14 words
A method as claimed in claim 13, wherein the protein is hGH or TNF.
References Cited (Backward Citations)
No citation records indexed.
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