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 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.
Thomas
L M
LM
Victorian Institute of Animal Science, Attwood, Australia.
Huntington
P J
PJ
Mead
L J
LJ
Wingate
D L
DL
Rogerson
B A
BA
Lew
A M
AM
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D015967:Gene Expression Regulation, Viral; D005982:Glutathione Transferase; D006736:Horses; D004860:Infectious Anemia Virus, Equine / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D015183:Restriction Mapping; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics*
D000956:Antigens, Viral; D004279:DNA, Viral; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C066797:transmembrane envelope glycoprotein gp45, retrovirus; D005982:Glutathione Transferase