Development of a simple typing method for the ovine Toll-like receptor 4 gene

Toll-like receptor 4 (TLR4) recognizes lipopolysaccharide (LPS) from Gram-negative bacteria, as well as a number of other ligands. Genetic variation in the TLR gene has been associated with altered immune responses to pathogens and results in variation in disease susceptibility. The objective of this work was to develop a simple and rapid genotyping system for ovine TLR4 and of a sensitivity that would allow detection of allelic variation in this gene. While variation in exon 3 of the ovine TLR4 gene has been described previously, we describe here an improved genotyping method. This method could not only reveal the four alleles that have been reported previously, but also revealed a further three new alleles of this gene in a population of 1670 New Zealand sheep. This improved genotyping method will be useful in understanding innate immune responses in individual sheep and could also be a useful tool for large-scale immune studies in sheep production systems.