A soluble recombinant fusion protein of the transmembrane envelope protein of equine infectious anaemia virus for ELISA

The use of the bacterial expression vector, pGex, to produce an abundant, soluble fusion protein of gp45 from equine infectious anaemia virus is described. Purification of the recombinant protein was achieved by one step affinity chromatography on immobilized glutathione using competitive elution so...

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Veröffentlicht in:	Veterinary microbiology 1992-06, Vol.31 (2), p.127-137
Hauptverfasser:	Thomas, Lynda M., Huntington, Peter J., Mead, Leeanne J., Wingate, Dianne L., Rogerson, Bruce A., Lew, Andrew M.
Format:	Artikel
Sprache:	eng
Schlagworte:	Amino Acid Sequence anemia infecciosa equina anemie infectieuse du cheval Animals antigene antigenos antigens Antigens, Viral - biosynthesis Antigens, Viral - genetics Base Sequence Biological and medical sciences Blotting, Western caballos cheval Chromatography, Affinity Cloning, Molecular DNA, Viral - chemistry elisa Enzyme-Linked Immunosorbent Assay equine infectious anaemia equine infectious anemia virus Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Viral Glutathione Transferase Horses Infectious Anemia Virus, Equine - chemistry Infectious Anemia Virus, Equine - genetics Microbiology Molecular Sequence Data Oligonucleotides - chemistry Polymerase Chain Reaction proteinas proteine proteins Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - genetics Restriction Mapping Techniques used in virology test elisa Viral Envelope Proteins - chemistry Viral Envelope Proteins - genetics Virology
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Zusammenfassung:	The use of the bacterial expression vector, pGex, to produce an abundant, soluble fusion protein of gp45 from equine infectious anaemia virus is described. Purification of the recombinant protein was achieved by one step affinity chromatography on immobilized glutathione using competitive elution so no harsh conditions were required. This provides a readily available antigen that is defined, plentiful and cheap. Yields of 3.5 mg of purified soluble protein/litre of bacterial culture were obtained. This antigen was found to be suitable for ELISA. Background reactivity to either the glutathione-S-transferase (GST) fusion partner by immune sera or the EIA-GST fusion protein by normal sera were negligible.
ISSN:	0378-1135 1873-2542
DOI:	10.1016/0378-1135(92)90071-Z