Isolation and in vitro translation of messenger RNA encoding allergens of the cat flea, Ctenocephalides felis.
Polyadenylated [poly(A)+] mRNA was isolated from the cat flea, Ctenocephalides felis by oligothymidylic acid-cellulose spin column chromatography and translated in vitro using a cell-free rabbit reticulocyte system. The relative incorporation of 35S-methionine into trichloroacetic acid-precipitable translation products obtained using poly (A+) mRNA was 48.5-fold over control translations performed without added mRNA. SDS-PAGE analysis of the translation products in combination with autoradiography showed that many proteins with apparent molecular weights in the range 14-90 K were synthesized. Immunoprecipitation studies performed using flea-allergic dog sera showed that several of the synthesized proteins corresponded with flea allergens. The allergenicity of the lysates was also confirmed by skin testing.
Greene
W K
WK
Institute for Molecular Genetics and Animal Disease, School of Veterinary Studies, Murdoch University, Australia.
Penhale
W J
WJ
Thompson
R C
RC
D000485:Allergens / Q000235:genetics*; D000818:Animals; D002415:Cats; D004283:Dog Diseases / Q000276:immunology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D006967:Hypersensitivity / Q000276:immunology / Q000662:veterinary; D066298:In Vitro Techniques; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D005423:Siphonaptera / Q000276:immunology*
D000485:Allergens; D012333:RNA, Messenger; D011061:Poly A