Production of a monospecific antiserum against the early region 1A proteins of adenovirus 12 and adenovirus 5 by an adenovirus 12 early region 1A-beta-galactosidase fusion protein antigen expressed in bacteria.
Antisera were prepared against the amino acid sequences encoded within the N-terminal half of the adenovirus 12 (Ad12) early region 1A (E1A) gene. This was accomplished by construction of a plasmid vector which encoded the N-terminal 131 amino acids of Ad12 E1A joined in frame to the coding sequence of beta-galactosidase. After induced synthesis in Escherichia coli, the Ad12 E1A-beta-galactosidase fusion protein (12-1A-FP) was extracted with urea and used to raise antibodies in rabbits. The 12-1A-FP antisera immunoprecipitated major phosphoproteins of 39,000 and 37,000 apparent molecular weights from Ad12-transformed and infected cells. The 12-1A-FP antisera also immunoprecipitated E1A phosphoproteins from Ad5-transformed and infected cells. Immunospecificity of the 12-1A-FP antisera was demonstrated by the ability of 12-1A-FP antigen to block immunoprecipitation of E1A proteins. Furthermore, E1A proteins immunoprecipitated from in vivo-labeled cells comigrated with those translated in vitro by RNA that had been hybridization selected to E1A DNA.
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005696:Galactosidases / Q000235:genetics*; D007106:Immune Sera; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D001616:beta-Galactosidase / Q000235:genetics*
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D007106:Immune Sera; D004262:DNA Restriction Enzymes; D005696:Galactosidases; D001616:beta-Galactosidase