Allelic variation of the ovine Toll-like receptor 4 gene

The family of Toll-like receptors (TLRs) is responsible for recognition of pathogen-associated molecular patterns (PAMPs), and Toll-like receptor 4 (TLR4) recognizes not only exogenous ligands such as the lipopolysaccharide (LPS) of Gram-negative bacteria, but also a number of endogenous ligands. Variation in the nucleotide sequence of the ovine TLR4 gene was investigated by amplification of a fragment containing a putative ligand-binding region using polymerase chain reaction (PCR), followed by single-strand conformational polymorphism (PCR-SSCP) analysis and DNA sequencing. Four novel SSCP patterns, representing four different sequences, were identified. Either one or two different sequences were detected in individual sheep and all the sequences identified shared high homology to the TLR4 sequences from a variety of species, suggesting that these sequences represent allelic variants of the ovine TLR4 gene. Fourteen single nucleotide polymorphisms (SNPs) were detected, and 79% (11 of 14) of SNPs were non-synonymous substitutions that would result in amino acid changes. Variation detected here might have an impact on pattern recognition and hence affect the immune response to pathogens.