Isolation of monoclonal antibodies specific for Rous sarcoma virus structural, polymerase and transforming proteins and their use for the study of mutant virus-infected cells.
Monoclonal antibodies were developed that are specific for Rous sarcoma virus structural, polymerase (reverse transcriptase) and transforming proteins. The monoclonal antibodies were shown to bind to purified virus proteins in an indirect 125I-labelled Protein A binding assay suitable for screening even very large numbers of hybridomas. Additional tests for specificity included radioimmunoprecipitation of purified virus structural proteins P12 and P27, of reverse transcriptase subunits alpha and beta, and of the transforming protein pp60v-src. Pilot immunofluorescence and protein kinase assays of the expression of virus proteins in avian and mammalian cells infected by wild-type virus as well as by temperature-sensitive, transformation-defective virus mutants revealed that synthesis of virus structural and transforming proteins is hardly affected by changes in temperature, whereas the pp60v-src-associated kinase activity is temperature-sensitive in cells infected by most, but not all the virus mutants.
Kurth
R
R
Tanaka
T
T
Moelling
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000096:biosynthesis; D000952:Antigens, Polyomavirus Transforming; D000956:Antigens, Viral / Q000276:immunology; D000957:Antigens, Viral, Tumor / Q000276:immunology*; D001358:Avian Sarcoma Viruses / Q000276:immunology*; D002642:Chick Embryo; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D051379:Mice; D009154:Mutation; D019384:Nucleic Acid Synthesis Inhibitors; D012194:RNA-Directed DNA Polymerase / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*; D015678:Viral Structural Proteins
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000952:Antigens, Polyomavirus Transforming; D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D019384:Nucleic Acid Synthesis Inhibitors; D014764:Viral Proteins; D015678:Viral Structural Proteins; D012194:RNA-Directed DNA Polymerase