Nucleotide sequence of the porcine p53 cDNA, and the detection of recombinant porcine p53 expressed in vitro with a variety of anti-p53 antibodies.
The cDNA of porcine p53 was cloned and sequenced by a reverse transcriptase polymerase chain reaction (RT - PCR) approach with primers based on regions of homology between all known p53 sequences. The p53 cDNA was found to be 87% conserved to human p53 and 86% conserved to bovine p53 at the nucleotide level. The porcine p53 sequence was inserted into an expression vector and recombinant protein expressed in vitro. An approximately 50 kDa protein was detected by Western blotting using both polyclonal and monoclonal anti-p53 antibodies. The sequence data of porcine p53 and the ability to detect expressed protein with various anti-p53 antibodies will allow the p53 status of the pig population, and the role of p53 in porcine tumours, to be assessed. An understanding of tumour development in the pig may be important if pig cells, tissues or organs are to be used in the treatment of humans.
Burr
P D
PD
Department of Veterinary Clinical Studies, University of Glasgow, Glasgow, G61 1QH.
Argyle
D J
DJ
Reid
S W
SW
Nasir
L
L
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics / Q000276:immunology; D003429:Cross Reactions; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D016158:Genes, p53; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D016415:Sequence Alignment; D013552:Swine / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53