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ES2096529A1 - Process for the preparation and purification of the protein coded by ORF-5 in porcine reproductive and respiratory syndrome virus by affinity chromatography - Google Patents

Process for the preparation and purification of the protein coded by ORF-5 in porcine reproductive and respiratory syndrome virus by affinity chromatography

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Publication number
ES2096529A1
ES2096529A1 ES09500807A ES9500807A ES2096529A1 ES 2096529 A1 ES2096529 A1 ES 2096529A1 ES 09500807 A ES09500807 A ES 09500807A ES 9500807 A ES9500807 A ES 9500807A ES 2096529 A1 ES2096529 A1 ES 2096529A1
Authority
ES
Spain
Prior art keywords
orf
protein
respiratory syndrome
porcine reproductive
syndrome virus
Prior art date
Legal status (The legal status 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 status listed.)
Granted
Application number
ES09500807A
Other languages
Spanish (es)
Other versions
ES2096529B1 (en
Inventor
Arganda Jose Maria Castro
Ferreiro Paloma Suarez
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universidad Complutense de Madrid
Original Assignee
Universidad Complutense de Madrid
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 Universidad Complutense de Madrid filed Critical Universidad Complutense de Madrid
Priority to ES9500807A priority Critical patent/ES2096529B1/en
Publication of ES2096529A1 publication Critical patent/ES2096529A1/en
Application granted granted Critical
Publication of ES2096529B1 publication Critical patent/ES2096529B1/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Process for the preparation and purification of the protein coded by ORF-5 in porcine reproductive and respiratory syndrome virus by affinity chromatography The protein coded by ORF-5 of porcine reproductive and respiratory syndrome virus is prepared and purified by its expression in eukaryotic cells. In order to achieve this the coded sequence of the said protein is obtained by PCR, cloned in pGEM-T vector, subcloned in pRSET vector which provides a histidine tag for the sequence prior to the coding sequence, and again subcloned in expression vector pTM-1 for expression as a fusion protein, serving in turn as a basis for the synthesis of a recombinant vaccinia virus for use using double infection protocols with a vaccinia virus which coagulates T7 phage polymerase in eukaryotic cells. The protein expressed in this way is purified by combining it with a nickel acetonitrile resin as a result of the affinity of the histidines for this metal.
ES9500807A 1995-04-26 1995-04-26 PROCEDURE FOR THE PREPARATION AND PURIFICATION BY AFFINITY CHROMATOGRAPHY OF PROTEIN CODED BY ORF-5 OF THE VIRUS OF THE REPRODUCTIVE AND RESPIRATORY SWINE SYNDROME. Expired - Fee Related ES2096529B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
ES9500807A ES2096529B1 (en) 1995-04-26 1995-04-26 PROCEDURE FOR THE PREPARATION AND PURIFICATION BY AFFINITY CHROMATOGRAPHY OF PROTEIN CODED BY ORF-5 OF THE VIRUS OF THE REPRODUCTIVE AND RESPIRATORY SWINE SYNDROME.

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES9500807A ES2096529B1 (en) 1995-04-26 1995-04-26 PROCEDURE FOR THE PREPARATION AND PURIFICATION BY AFFINITY CHROMATOGRAPHY OF PROTEIN CODED BY ORF-5 OF THE VIRUS OF THE REPRODUCTIVE AND RESPIRATORY SWINE SYNDROME.

Publications (2)

Publication Number Publication Date
ES2096529A1 true ES2096529A1 (en) 1997-03-01
ES2096529B1 ES2096529B1 (en) 1997-11-16

Family

ID=8290189

Family Applications (1)

Application Number Title Priority Date Filing Date
ES9500807A Expired - Fee Related ES2096529B1 (en) 1995-04-26 1995-04-26 PROCEDURE FOR THE PREPARATION AND PURIFICATION BY AFFINITY CHROMATOGRAPHY OF PROTEIN CODED BY ORF-5 OF THE VIRUS OF THE REPRODUCTIVE AND RESPIRATORY SWINE SYNDROME.

Country Status (1)

Country Link
ES (1) ES2096529B1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999037784A1 (en) * 1998-01-27 1999-07-29 Her Majesty In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada Porcine reproductive and respiratory syndrome oral vaccine production in plants

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5047513A (en) * 1986-07-10 1991-09-10 Hoffmann-La Roche Inc. Metal chelate resins
US5310663A (en) * 1987-03-10 1994-05-10 Hoffman-La Roche Inc. Affinity peptides

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5047513A (en) * 1986-07-10 1991-09-10 Hoffmann-La Roche Inc. Metal chelate resins
US5310663A (en) * 1987-03-10 1994-05-10 Hoffman-La Roche Inc. Affinity peptides

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LÍNNERDAL, B. et al., Metal chelate affinity chromatography of proteins, Journal of Applied Biochemistry, 1982, Vol. 4, pßg. 203-208. *Ver todo el documento* *
MEULENBERG J.J.M. et al., Lelystad virus, the causative agent of procine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV, Virology, 1993, Vol.192, pßg.62-72. *Ver todo el documento* *
PORATH, J. et al., Immobilized metal ion affinity adsorption and immobilized metal ion affinity chromatography of biomaterials. Serum protein affinities for gel-immobilized iron and nickel ions. Biochemistry, 1983, Vol.22, pßg.1621-1630 *Ver todo el documento* *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999037784A1 (en) * 1998-01-27 1999-07-29 Her Majesty In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada Porcine reproductive and respiratory syndrome oral vaccine production in plants

Also Published As

Publication number Publication date
ES2096529B1 (en) 1997-11-16

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