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Comparison of the in vitro translation capacity ofTaenia crassicepsmetacestode mRNA prepared by the phenol and cesium chloride method

✍ Scribed by R. Kleine-Herzbruch; E. Geyer

Book ID
104794714
Publisher
Springer-Verlag
Year
1988
Tongue
English
Weight
835 KB
Volume
74
Category
Article
ISSN
1432-1955

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✦ Synopsis

Total RNA of T. crassiceps metacestodes harvested from male and female NMRI mice was prepared by both the phenol extraction technique and cesium chloride (CsCl) gradient centrifugation. mRNA was selected by oligo (dT)-cellulose affinity chromatography and used as the template for the in vitro translation of parasite polypeptides in a cell-free rabbit reticulocyte lysate. The template activity of the mRNA obtained after CsCl preparation was clearly higher, as shown by the amount of 35S-methionine incorporated into the translation products and by fluorographed SDS-PAGE of the synthesized labelled polypeptides. SDS-PAGE fluorographs of antigens encoded by the mRNA prepared by CsCl centrifugation and selected by immunoprecipitation using purified IgG antibodies of T. crassiceps-infected mice (day 80 postinfection) exhibited seven labelled polypeptides of about 65, 46, 45, 42, 34, 29 kDa and a predominant 20-kDa antigen. The latter polypeptide was the only one recognized by the antibodies amongst the in vitro translation products directed by mRNA prepared by the phenol method.

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✍ PRIOR, RONALD L. πŸ“‚ Article πŸ“… 2001 πŸ› Elsevier 🌐 English βš– 986 KB

This work contains over thirty chapters by leading researchers in the field of oxidative biology, originally presented as articles in an extended Forum in the highly-cited journal, *Free Radical Biology & Medicine*. The papers in this Forum (or Symposium-in-print) spanned seven issues of the journal