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An antibody to a synthetic peptide that recognises SV40 small‐t antigen.
Author(s) -
Harvey R.,
Faulkes R.,
Gillett P.,
Lindsay N.,
Paucha E.,
Bradbury A.,
Smith A.E.
Publication year - 1982
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1982.tb01193.x
Subject(s) - biology , peptide , antigen , antibody , virology , peptide sequence , microbiology and biotechnology , biochemistry , immunology , gene
A peptide Tyr.Arg.Asp.Leu.Lys.Leu corresponding to the carboxy‐terminal six amino acids of small‐t antigen predicted from the DNA sequence of SV40 was synthesised, coupled to bovine serum albumin and to ovalbumin and used to raise antibody in rabbits. The sera obtained immunoprecipitated [125I]peptide. It also recognised SV40 small‐t that was synthesised in vitro from SV40 mRNA or extracted from SV40 infected monkey cells. The immunoprecipitation of small‐t was inhibited by added peptide. To demonstrate that the determinant was present at the carboxy‐terminal end of the molecule, truncated versions of small‐t coded for by 0.54‐0.59 deletion mutants were tested. dl 890 small‐t, which contains an in‐phase deletion removing nine amino acids but leaving the carboxy‐terminal sequences intact, was recognised by the antipeptide serum. By contrast dl 885 small‐t, which has an out‐of‐phase deletion leading to an altered carboxy terminus coded in an alternative reading frame, was not recognised. The data confirm the location and specificity of the determinant recognised on small‐t by the antipeptide serum.