Use of proteinase K in the excystation of Sarcocystis cruzi sporocysts for in vitro culture and DNA extraction.
Proteinase K was used for the cleaning of Sarcocystis cruzi (Apicomplexa) sporocysts prior to excystation. Bovine pulmonary endothelial cell cultures inoculated with the excysted sporozoites remained free of bacterial contamination for the duration of the experiment and had high yields of merozoites. The excysted sporozoites also yielded genomic DNA that could be labelled efficiently with 32P dATP by the random priming method.
Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, Canada.
W
W
Ndiritu
R J
RJ
Cawthorn
F S
FS
Kibenge
D000818:Animals; D002417:Cattle; D002460:Cell Line; D016054:DNA, Protozoan / Q000302:isolation & purification*; D019286:Endopeptidase K; D004727:Endothelium / Q000166:cytology / Q000469:parasitology; D008168:Lung / Q000166:cytology / Q000469:parasitology; D012522:Sarcocystis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D012697:Serine Endopeptidases / Q000378:metabolism*
D016054:DNA, Protozoan; D012697:Serine Endopeptidases; D019286:Endopeptidase K