Preparation of Poliovirus Labeled with Phosphorus-33
AUTOR(ES)
Sobsey, Mark D.
RESUMO
Phosphorus-33 (33P), a weak (0.25 Mev) beta-emitting isotope of phosphorus with a half-life of 25 days, has been used to label poliovirus in cell culture. HeLa cell monolayers were depleted of phosphate and then labeled by incubating at 37 C in a medium (LM) containing about 10 μCi of 33P as orthophosphate per ml. Labeled cells were infected at a high multiplicity with poliovirus type 1 and incubated for 8 hr in LM medium. Virus from infected cells was then concentrated and purified. Virus purity was confirmed by comparison of virus infectivity and radioactivity after CsCl density gradient centrifugation and by observing purified virus preparations with electron microscopy. With the method described, yields of about 1010 to 5 × 1010 plaque-forming units (PFU) of highly purified poliovirus with specific activities of about 3 × 10-4 to 10-3 disintegrations per min per PFU have been obtained from 1.5 × 108 to 3.0 × 108 HeLa cells.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=380397Documentos Relacionados
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