EP1106697A1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Mass secretion/expression system of true mk family protein
Assignee: Meiji Dairies Corp
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Filing: 1999-08-10
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Granted: —
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Expires: 2019-08-10
Abstract
Abstract Under the regulation of a methanol-inducible alcohol oxidase gene (AOX1) promoter and a transcription termination sequence originating from Pichia pastor is, Pichia yeast, which has been transformed with an expression vector containing an MK family protein gene ligated to an alpha 1 factor signal sequence originating from Saccharomyces cerevisiae, is cultured and an intact MK family protein is thus expressed and secreted in a large amount into the culture supernatant.
Inventors
- 👤 Akira-Meiji Institute of Health Science MURASUGI
- 👤 Yukio-Meiji Institute of Health Science ASAMI
- 👤 Isao-Meiji Milk Products Co. Ltd. KIDO
- 👤 Hideshi-Meiji Milk Products Co. Ltd. KUMAI
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 06:01
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/79; C12N15/80; C12N15/81; C07K2319/00; C07K2319/01; C07K2319/02
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
37 words
A vector for secretory expression of an intact MK family protein
by methylotrophic yeast, said vector comprising a gene encoding
a mature MK family protein ligated to a signal sequence of α1 factor
derived from Saccharomyces cerevisiae.
Claim 2
Dependent Claim
111 words
The vector according to claim 1 comprising components (a) to (g)
below: (a) a promoter sequence of a methanol-inducible alcohol oxidase
gene (AOX1) derived from Pichia pastoris, (b) a signal sequence of α1 factor derived from Saccharomyces
cerevisiae , (c) a gene encoding a mature MK family protein, wherein said gene
is ligated to (b), (d) a transcription termination sequence of a methanol-inducible
alcohol oxidase gene (AOX1) derived from Pichia pastoris , (e) a selection marker gene functioning in Escherichia coli and
methylotrophic yeast, (f) a replication origin functioning in Escherichia coli , and (g) 5' AOX1 and 3' AOX1 for the site-specific homologous
recombination to a methylotrophic yeast chromosomal DNA.
Claim 3
Dependent Claim
14 words
The vector according to claim 1, wherein said MK family protein
is MK protein.
Claim 4
Dependent Claim
14 words
The vector according to claim 1, wherein said MK family protein
is PTN protein.
Claim 5
Dependent Claim
18 words
A transformant comprising methylotrophic yeast transformed with
the vector according to any one of claims 1 to 4.
Claim 6
Dependent Claim
27 words
The transformant according to claim 5, wherein said transformant
is pPIC9DP-hMK/SMD1168, said vector is the one according to claim
3, and said methylotrophic yeast is strain SMD1168.
Claim 7
Dependent Claim
26 words
The transformant according to claim 5, wherein said transformant
is pPIC9-hPTN/GS115, said vector is the one according to claim 4,
and methylotrophic yeast is strain GS115.
Claim 8
Independent Claim
29 words
A method for producing an intact MK family protein, said method
comprising culturing the transformant according to any one of claims
5 to 7 and recovering secretory expression products.
Claim 9
Dependent Claim
44 words
The method according to claim 8, said method comprising: (a) culturing the transformant according to claim 6, (b) inducing the expression of MK protein under the conditions of
20°C and pH 3 after the proliferation at pH 4, and (c) recovering secretory expression products.
References Cited (Backward Citations)
No citation records indexed.
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