Abstract
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.
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- HENRY C., YOUNGNER J. S. STUDIES ON THE STRUCTURE AND REPLICATION OF THE NUCLEIC ACID OF POLIOVIRUS. Virology. 1963 Oct;21:162–173. doi: 10.1016/0042-6822(63)90253-8. [DOI] [PubMed] [Google Scholar]
- HOYER B. H., BOLTON E. T., RITTER D. B., RIBI E. Simple method for preparation of purified radioactive poliovirus particles: electron micrograph. Virology. 1959 Apr;7(4):462–464. doi: 10.1016/0042-6822(59)90074-1. [DOI] [PubMed] [Google Scholar]
- Krisch R. E. Comparison of the lethal effects of 32P and 33P decay in E. coli and in bacteriophages. Int J Radiat Biol Relat Stud Phys Chem Med. 1970;18(3):259–266. doi: 10.1080/09553007014551071. [DOI] [PubMed] [Google Scholar]
- Phillips B. A., Summers D. F., Maizel J. V., Jr In vitro assembly of poliovirus-related particles. Virology. 1968 Jun;35(2):216–226. doi: 10.1016/0042-6822(68)90262-6. [DOI] [PubMed] [Google Scholar]
- Robinson J. R. 33P: a superior radiotracer for phosphorus? Int J Appl Radiat Isot. 1969 Jul;20(7):531–540. doi: 10.1016/0020-708x(69)90007-6. [DOI] [PubMed] [Google Scholar]
- TAYLOR J., GRAHAM A. F. Purification of poliovirus labeled with radiophosphorus. Virology. 1958 Oct;6(2):488–498. doi: 10.1016/0042-6822(58)90096-5. [DOI] [PubMed] [Google Scholar]