Cloning of cervine interferon-gamma cDNA by polymerase chain reaction

Abstract

As the first step in the development of a cervine IFN-γ assay for the diagnosis of tuberculosis in deer, cervine IFN-γ cDNA was amplified by polymerase chain reaction using primers based on the bovine IFN-γ sequence. A high level of amino acid homology was found between the cervine and the ovine and bovine sequences (94% and 91% respectively). There was less identity with the porcine, human, mouse and rat sequences (78%, 62%, 37% and 39%, respectively). The amino terminus of the mature IFN-γ protein, which is critical for interaction with its receptor and for triggering biological activity, is highly conserved between the cervine, bovine and ovine proteins. A monoclonal antibody-based sandwich enzyme immunoassay (EIA) specific for bovine IFN-γ also detects ovine but not cervine IFN-γ. This suggests that the antibodies recognise epitopes common to the bovine and ovine protein but not cervine IFN-γ. Seven amino acid residues that were common to the bovine and ovine sequence differed in the cervine sequence, suggesting that the specificity of the monoclonal antibodies may be dependent on one or more of these residues. The possibility of the development of an EIA for cervine IFN-γ as a commercial in vitro diagnostic assay for tuberculosis in deer is discussed.