Analysis of mRNA coding for blood-stage antigens of a rodent malarial parasite, Plasmodium yoelii: mRNA coding for a possible protective antigen purify as poly A-.
We have analyzed mRNA coding for blood-stage antigens of Plasmodium yoelii by using cellfree translation of poly A+ and poly A- RNA in conjunction with immunoprecipitations. Most of the antigens recognized by mouse hyperimmune serum to P. yoelii were coded by poly A+ mRNA ranging in size from 15S to 28S. However, certain P. yoelii antigens, notably those with m.w. greater than 150 kilodaltons (kd), were coded by mRNA that purified as being poly A-. Antigens recognized by a protective monoclonal antibody (McAb) were coded by such operationally poly A- RNA. Three polypeptides apparently coded by different poly A- RNA were immunoprecipitated by this McAb. With the use of another McAb, a poly A+ mRNA of about 19S was identified as coding for a polypeptide of 46 kd synthesized in cellfree translation reactions. The same McAb recognized a 34 kd polypeptide in metabolically labeled polypeptides of P. yoelii. This antigen appeared to be processed in vivo but not in vitro. The observation that some mRNA of P. yoelii purify as being poly A- has significant implications for the construction of cDNA libraries that employ poly A+ mRNA of malarial parasites: if it applies to other species of plasmodia, some potentially important operationally poly A- mRNA may not be represented in such libraries.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D005815:Genetic Code; D008288:Malaria / Q000097:blood / Q000276:immunology / Q000469:parasitology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010961:Plasmodium / Q000235:genetics / Q000276:immunology*; D011061:Poly A / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000941:Antigens; D012333:RNA, Messenger; D011061:Poly A