A simplified method for typing DRB alleles in deer
E.M. Linscott, C.M. Mackintosh, J.F.T. Griffin and A.M.
Crawford
AgResearch Molecular Biology Unit, Biochemistry Building, Otago
University, Cumberland Street, Dunedin
Proceedings of the New Zealand Society of Animal Production. 1998,
58: 13-15
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-labeled 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.
Keywords: NZSAPAB;
Major histocompatibility complex, Deer, DRB gene typing
Last Updated 12-09-1998
