WO1994018331A3 - Fusion proteins including a cleavage site recognized by a plant virus protease - Google Patents

Fusion proteins including a cleavage site recognized by a plant virus protease Download PDF

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Publication number
WO1994018331A3
WO1994018331A3 PCT/US1994/001176 US9401176W WO9418331A3 WO 1994018331 A3 WO1994018331 A3 WO 1994018331A3 US 9401176 W US9401176 W US 9401176W WO 9418331 A3 WO9418331 A3 WO 9418331A3
Authority
WO
WIPO (PCT)
Prior art keywords
plant virus
fusion proteins
cleavage site
proteins including
site recognized
Prior art date
Application number
PCT/US1994/001176
Other languages
French (fr)
Other versions
WO1994018331A2 (en
Inventor
Stephen A Johnston
William G Dougherty
Original Assignee
Univ Texas
Univ Oregon
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Univ Texas, Univ Oregon filed Critical Univ Texas
Priority to AT94907952T priority Critical patent/ATE294870T1/en
Priority to DE69434358T priority patent/DE69434358D1/en
Priority to AU61322/94A priority patent/AU6132294A/en
Priority to EP94907952A priority patent/EP0682709B1/en
Publication of WO1994018331A2 publication Critical patent/WO1994018331A2/en
Publication of WO1994018331A3 publication Critical patent/WO1994018331A3/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/23—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a GST-tag

Abstract

Methods for isolation and purification of recombinant proteins are described. Fusion proteins incorporating a cleavage site sensitive to proteolysis by a plant virus proteinase may be cleaved from carrier proteins to provide high yields of protein product. Methods employing a plant virus proteinase to cleave expressed fusion proteins are particularly suitable for obtaining proteolytically sensitive polypeptides in the presence of added cell protease inhibitors. Also disclosed are recombinant vectors useful for overproducing plant virus proteinases in a suitable host.
PCT/US1994/001176 1993-02-12 1994-01-31 Fusion proteins including a cleavage site recognized by a plant virus protease WO1994018331A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AT94907952T ATE294870T1 (en) 1993-02-12 1994-01-31 METHOD FOR PRODUCING PROTEINASE NLA FROM TOBACCO
DE69434358T DE69434358D1 (en) 1993-02-12 1994-01-31 METHOD OF PREPARING PROTEINASE NLA FROM TOBACCO
AU61322/94A AU6132294A (en) 1993-02-12 1994-01-31 Fusion proteins including a cleavage site recognized by a plant virus protease
EP94907952A EP0682709B1 (en) 1993-02-12 1994-01-31 Method for producing tobacco proteinase nla

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/021,603 US5532142A (en) 1993-02-12 1993-02-12 Method of isolation and purification of fusion polypeptides
US08/021,603 1993-02-12

Publications (2)

Publication Number Publication Date
WO1994018331A2 WO1994018331A2 (en) 1994-08-18
WO1994018331A3 true WO1994018331A3 (en) 1994-10-13

Family

ID=21805141

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1994/001176 WO1994018331A2 (en) 1993-02-12 1994-01-31 Fusion proteins including a cleavage site recognized by a plant virus protease

Country Status (6)

Country Link
US (1) US5532142A (en)
EP (1) EP0682709B1 (en)
AT (1) ATE294870T1 (en)
AU (1) AU6132294A (en)
DE (1) DE69434358D1 (en)
WO (1) WO1994018331A2 (en)

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Also Published As

Publication number Publication date
WO1994018331A2 (en) 1994-08-18
DE69434358D1 (en) 2005-06-09
EP0682709A1 (en) 1995-11-22
US5532142A (en) 1996-07-02
ATE294870T1 (en) 2005-05-15
AU6132294A (en) 1994-08-29
EP0682709B1 (en) 2005-05-04

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