PRODUCTION OF A POLYCLONAL ANTIBODY RAISED AGAINST RECOMBINANT FLOUNDER P53 PROTEIN

The cDNA encoding the wild type p53 protein from flounder, Platichthys flesus, was expressed in Escherichia coli using the GST fusion protei n system. Several milligrams of recombinant p53 protein were purified. This protein displayed an apparent molecular weight of 45000 Da, a va lue which is very similar to Xenopus p53, but significantly greater th an was expected based on the length of the open reading frame. Immuniz ation of rabbits against purified p53 protein allowed the production o f high titre polyclonal antiserum. This new polyclonal antibody recogn ized recombinant flounder p53 protein in Western blot. Cross reaction was also observed with recombinant Xenopus p53 protein but not with hu man p53 protein. Immunoblotting of the total protein extract from norm al flounder ovaries did not reveal any p53 expression. (C) 1998 Elsevi er Science Inc. All rights reserved.