The method of superlong sequencing to determine the nucleotide sequences of the cattle TLR genes
A representative set of nine modern cattle breeds from Russia were screened for the variability in genes coding for Toll-like receptors (TLRs). The variability in TLRs is, among other, a potential basis for reducing the risk of miscarriages due to infections in the early reproduction via breeding. The overlapping exon amplicons from genomic DNA samples of 575 animals were sequenced using the PacBio platform. 64 putative mutations in the coding sequences were identified with a considerable proportion of novel SNPs. The structural variants were annotated according to their functional significance. Further on, 31 of the structural variants were individually genotyped using the primer extension method. As a result of the research, gene variability of this group of TLR genes was fully characterized in the representative cattle breed panel of Russia. The information will be used to explore the associations of the found TLR variants with the reproductive traits like pregnancy that might be affected by intrauterine infections and general infections of cattle.
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