Cell free synthesis of Toxoplasma gondii antigens.
Functionally active poly(A)-containing mRNA was isolated from tachyzoites of the RH strain of Toxoplasma gondii. The T. gondii mRNA was capable of directing the synthesis of proteins in a wheat germ in vitro translation system, but not in a cell free system derived from rabbit reticulocyte lysate. Efficient translation in the wheat germ system required spermine and exogenous tRNA. Amino acid incorporation was maximal at 110 mM K+ and 1.8 mM Mg2+. Tachyzoite antigens synthesized in vitro were immunoprecipitated with T. gondii antibodies from rabbits, mice and humans. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitated polypeptides yielded patterns that differed according to antibody source, but all T. gondii antibody preparations reacted with a translation product with an apparent molecular weight of 24 000.
Prince
J B
JB
Koven-Quinn
M A
MA
Remington
J S
JS
Sharma
S D
SD
D000818:Animals; D000941:Antigens / Q000235:genetics* / Q000302:isolation & purification; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008970:Molecular Weight; D010944:Plants / Q000378:metabolism; D011061:Poly A / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics*; D014908:Triticum / Q000378:metabolism
D000941:Antigens; D012333:RNA, Messenger; D011061:Poly A