Johnson A M, McDonald P J, Illana S
Mol Biochem Parasitol. 1986 Mar;18(3):313-20. doi: 10.1016/0166-6851(86)90088-5.
RNA was extracted from purified tachyzoites of Toxoplasma gondii (RH strain) by sequential centrifugation in guanidine hydrochloride, urea, and lithium chloride. The subunits of the RNA were characterized by denaturing and non-denaturing electrophoresis in agarose gels. Poly(A)+-RNA, purified by oligo(dT)-cellulose affinity chromatography, was translated in a rabbit reticulocyte lysate assay and the products were immunoprecipitated with an experimentally infected mouse serum and a naturally infected human serum. After sodium dodecyl sulphate-polyacrylamide gel electrophoresis, fluorography of the polypeptides confirmed that the mRNA translated specific parasite antigens.
通过在盐酸胍、尿素和氯化锂中连续离心,从纯化的刚地弓形虫(RH株)速殖子中提取RNA。RNA的亚基通过琼脂糖凝胶中的变性和非变性电泳进行表征。通过寡聚(dT)-纤维素亲和色谱纯化的聚(A)+ -RNA在兔网织红细胞裂解物测定中进行翻译,产物用实验感染的小鼠血清和自然感染的人血清进行免疫沉淀。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后,多肽的荧光自显影片证实mRNA翻译了特定的寄生虫抗原。