Glycosylation of type-IV fimbriae of Dichelobacter nodosus

WOS: 000251924700017PubMed ID: 17681435Dichelobacter nodosus is the causative agent of ovine footrot and the type-IV fimbriae on this bacterium are essential for maintaining its virulence. In this study, we reveal that these fimbriae are glycosylated. This was demonstrated in several ways: by the detection of carbohydrate on fimbrial protein using periodic acid Schiff reagent (PAS) staining of SDS-PAGE gels and by demonstrating enzymatic deglycosylation and by analysis of the amino acid sequences derived from the fimA gene, whereby the gene from isolates of D. nodosus that appeared to be glycosylated had potential glycosylation sites both inside and outside of the variable region of fimA. The results would also explain the observation that the calculated molecular weight of fimA from some D. nodosus serotypes does not correlate with the apparent size determined from electrophoretic mobility. (C) 2007 Elsevier B.V. All rights reserved

Haplotypes and Sequence Variation in the Ovine Adiponectin Gene (ADIPOQ)

The adiponectin gene (ADIPOQ) plays an important role in energy homeostasis. In this study five separate regions (regions 1 to 5) of ovine ADIPOQ were analysed using PCR-SSCP. Four different PCR-SSCP patterns (A1-D1, A2-D2) were detected in region-1 and region-2, respectively, with seven and six SNPs being revealed. In region-3, three different patterns (A3-C3) and three SNPs were observed. Two patterns (A4-B4, A5-B5) and two and one SNPs were observed in region-4 and region-5, respectively. In total, nineteen SNPs were detected, with five of them in the coding region and two (c.46T/C and c.515G/A) putatively resulting in amino acid changes (p.Tyr16His and p.Lys172Arg). In region-1, -2 and -3 of 316 sheep from eight New Zealand breeds, variants A1, A2 and A3 were the most common, although variant frequencies differed in the eight breeds. Across region-1 and region-3, nine haplotypes were identified and haplotypes A1-A3, A1-C3, B1-A3 and B1-C3 were most common. These results indicate that the ADIPOQ gene is polymorphic and suggest that further analysis is required to see if the variation in the gene is associated with animal production traits

Variation in the Caprine KAP24-1 Gene Affects Cashmere Fibre Diameter

The keratin-associated proteins (KAPs) are structural components of cashmere fibres. The gene encoding the high-sulphur (HS)-KAP24-1 (KRTAP24-1) has been identified in humans and sheep, but it has not been described in goats. In this study, we report the identification of caprine KRTAP24-1, describe variation in this gene, and investigate the effect of this variation on cashmere traits. A search for sequences orthologous to the ovine gene in the goat genome revealed a 774 bp open reading frame on chromosome 1, which could encode an HS-KAP. Based on this goat genome sequence and comparison with ovine KRTAP24-1 sequences, polymerase chain reaction (PCR) primers were designed to amplify an 856 bp fragment that would contain the entire coding region of the putative caprine KRTAP24-1. Use of this PCR amplification with subsequent single-strand conformation polymorphism (SSCP) analysis of the amplicons identified four distinct patterns of DNA bands on gel electrophoresis, with these representing four different DNA sequences (A to D), in 340 Longdong cashmere goats reared in China. The variant sequences had the highest similarity to KRTAP24-1 sequences from sheep and humans, suggesting that they are variants of caprine KRTAP24-1. Nine single-nucleotide polymorphisms (SNPs) were detected in the gene, including four non-synonymous SNPs and an SNP in proximity to the ATG start codon. Of the three common genotypes (AA, AB, and BB) found in these Longdong cashmere goats, cashmere fibres from goats of genotype AA had lower mean fibre diameter (MFD) than did those of genotype AB, and cashmere fibres from goats of genotype AB had lower MFD than did those from goats of genotype BB

Polymorphism of ovine <i>FABP4</i> and sequence homology with other species.

<p>Five and four allele variants are respectively identified in region I (exon 2-intron 2) and region II (exon 3-intron 3) by (<b>a</b>) PCR-SSCP and (<b>b</b>) DNA sequencing <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088691#pone.0088691-Yan1" target="_blank">[11]</a>. The <i>FABP4</i> sequences from other species used to construct the homology trees (c) are: NC_007312 (cattle), EF061481 (pig), HQ384160 (human), ENSFCAG00000032028 (cat) and ENSMUSG00000062515 (mouse). The nucleotide numbering follows the nomenclature described on <a href="http://www.hgvs.org/mutnomen/" target="_blank">www.hgvs.org/mutnomen/</a>.</p

The presence of potential “recombination hotspots” in ovine <i>FABP4</i> and the haplotypes across two gene regions that were inherited from the seven NZ Romney sires.

<p>The locations of these two regions in ovine <i>FABP4</i> are indicated. An unfilled triangle represents uble overlapping <i>chi</i>-like sequences (GCTGGTGCTGGTGA), and a horizontal line filled triangle represents the <i>CRE</i>-like sequence (ATGAAGTCA). The CCTCCCT motifs and variants are indicated by dot filled triangles, and the CCAAT motifs are shown as filled triangles. The numbering of nucleotide positions follows the nomenclature described at <a href="http://www.hgvs.org/mutnomen/" target="_blank">www.hgvs.org/mutnomen/</a></p

Paternal haplotypes of ovine <i>FABP4</i> detected in individual sire-lines.

<p>* Recombined minor sire haplotypes.</p