Isolation and in vitro translation of messenger RNA encoding allergens of the cat flea, Ctenocephalides felis
Greene, W.K., Penhale, W.J. and Thompson, R.C.A. (1993) Isolation and in vitro translation of messenger RNA encoding allergens of the cat flea, Ctenocephalides felis. Veterinary Immunology and Immunopathology, 37 (1). pp. 15-23.
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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.
|Publication Type:||Journal Article|
|Murdoch Affiliation:||School of Veterinary Studies|
|Copyright:||© Elsevier B.V.|
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