ISOLATION AND INVITRO TRANSLATION OF MESSENGER-RNA ENCODING ALLERGENSOF THE CAT FLEA, CTENOCEPHALIDES-FELIS

Polyadenylated [poly(A)+] mRNA was isolated from the cat flea, Ctenoce phalides felis by oligothymidylic acid-cellulose spin column chromatog raphy and translated in vitro using a cell-free rabbit reticulocyte sy stem. The relative incorporation of S-35-methionine into trichloroacet ic acid-precipitable translation products obtained using poly (A+) mRN A was 48.5-fold over control translations performed without added mRNA . SDS-PAGE analysis of the translation products in combination with au toradiography showed that many proteins with apparent molecular weight s in the range 14-90 K were synthesized. Immunoprecipitation studies p erformed using flea-allergic dog sera showed that several of the synth esized proteins corresponded with flea allergens. The allergenicity of the lysates was also confirmed by skin testing.