Production of recombinant extracellular domains of canine desmoglein 1 (Dsg1) by baculovirus expression.
The aim of this study was to generate a recombinant protein to represent the entire extracellular domain of canine desmoglein 1 (Dsg1), a desmosomal cell-cell adhesion molecule, by the baculovirus expression system. Cotransfection of a baculovirus transfer vector containing the cDNA for the entire extracellular domain of canine Dsg1 with baculovirus DNA into insect cells resulted in the secretion of soluble canine Dsg1 into insect culture supernatants. Immunoreactivity of 11 human pemphigus foliaceus (PF) sera against the cell surface of canine keratinocytes was completely removed when the sera were preincubated with the canine Dsg1 baculoprotein. This recombinant canine Dsg1 produced by baculovirus shares the major epitopes of the authentic canine Dsg1 recognized by human PF sera, and will be useful in studying the molecular pathophysiological mechanisms in PF and impetigo in canine patients.
Nishifuji
Koji
K
Department of Veterinary Internal Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan. kojimail@sc.itc.keio.ac.jp
Amagai
Masayuki
M
Nishikawa
Takeji
T
Iwasaki
Toshiroh
T
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015820:Cadherins / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology; D051183:Desmoglein 1; D004283:Dog Diseases / Q000276:immunology; D004285:Dogs; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D015603:Keratinocytes / Q000276:immunology; D010392:Pemphigus / Q000276:immunology / Q000662:veterinary; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018411:Spodoptera / Q000821:virology
D015820:Cadherins; D051183:Desmoglein 1; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D012313:RNA