Abstract

Major histocompatibility complex (MHC) genes present foreign antigens to the immune system and polymorphism in these genes may have a key role in the variability of the host response to pathogenic challenge. The DRB genes are part of the MHC class II family whose proteins present foreign antigenic peptides on the surface of the macrophage for recognition by the CD4+T cells of the immune response. The genes are highly variable with over 100 different variants found in some species. The goal of this study was the development and testing of a simplified system for differentiating between the different DRB alleles already identified in red deer. The typing method chosen used DNA hybridisation to discriminate between the DRB alleles. The RNA from each deer was extracted from cultured leukocytes. Reverse transcription followed by PCR was used to amplify the DRB region. Amino-linked oligonucleotides representing variable regions of the DRB region were covalently bound to a nylon membrane and probed with end-labelled PCR product. The degree of binding of the labelled PCR products to the membrane-bound oligonucleotides was used to discriminate between the different DRB genes amplified. This proved to be a simple and robust method of typing which eliminated any requirement for electrophoresis.

EM, Linscott, CM Mackintosh, JFT Griffin, and AM Crawford

Proceedings of the New Zealand Society of Animal Production, Volume 58, , 13-15, 1998