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PMID: 196098 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Complementation rescue of Rous sarcoma virus from transformed mammalian cells by polyethylene glycol-mediated cell fusion.

Journal of virology ·Vol. 23 ·No. 1 ·1977-07-00 ·Pages 133-41

Steimer KS, Boettiger D

Abstract

Polyethylene glycol (PEG) is effective as a fusing agent for the rescue of virus from Rous sarcoma virus-transformed mammalian cells. The procedure of PEG-mediated rescue of virus from virogenic cell lines is described, and the technique is compared with that of Sendai virus-mediated rescue. Virus may be rescued quantitatively from virogenic cell lines by plating mitomycin C-killed transformed mammalian cells with chicken embryo cells, treating the monolayers with 50% PEG and overlaying the monolayers with focus agar. The number of foci that appeared reflected the number of heterokaryons in the fusion mixtures that released infectious virus. PEG gave reproducible results in virus rescue experiments with an efficiency equal to the best Sendai virus preparations. In addition to the description of the technique for PEG-mediated virus rescue from virogenic cell lines, a method for virus rescue from nonvirogenic lines is presented. Preinfection of the chicken embryo cells with helper avian leukosis virus (Rous-associated virus) prior to fusion with mammalian cells transformed by defective viruses complements the virus defect. We examined four nonvirogenic cell lines, and all released infectious virus in the complementation rescue assay.

MeSH Terms
Animals Avian Leukosis Virus/growth & development Avian Sarcoma Viruses/growth & development,isolation & purification Cell Fusion Cell Line Cell Transformation, Neoplastic Chickens Cricetinae Hybrid Cells Parainfluenza Virus 1, Human/growth & development Polyethylene Glycols/pharmacology Quail Rats Virus Replication
Chemicals
Polyethylene Glycols
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Steimer K S
Boettiger D
References (30)
30 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.23.1.133-141.1977
Published
1977-07-00
Pages
133-41
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC515809
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.23.1.133-141.1977 PMC Full Text View journal details
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