| 1. |
- Bellone, Rebecca R, et al.
(författare)
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Fine-mapping and mutation analysis of TRPM1 : a candidate gene for leopard complex (LP) spotting and congenital stationary night blindness in horses
- 2010
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Ingår i: Briefings in Functional Genomics & Proteomics. - : Oxford University Press (OUP). - 1473-9550 .- 1477-4062 .- 2041-2649 .- 2041-2657. ; 9:3, s. 193-207
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Tidskriftsartikel (refereegranskat)abstract
- Leopard Complex spotting occurs in several breeds of horses and is caused by an incompletely dominant allele (LP). Homozygosity for LP is also associated with congenital stationary night blindness (CSNB) in Appaloosa horses. Previously, LP was mapped to a 6 cm region on ECA1 containing the candidate gene TRPM1 (Transient Receptor Potential Cation Channel, Subfamily M, Member 1) and decreased expression of this gene, measured by qRT-PCR, was identified as the likely cause of both spotting and ocular phenotypes. This study describes investigations for a mutation causing or associated with the Leopard Complex and CSNB phenotype in horses. Re-sequencing of the gene and associated splice sites within the 105 624 bp genomic region of TRPM1 led to the discovery of 18 SNPs. Most of the SNPs did not have a predictive value for the presence of LP. However, one SNP (ECA1:108,249,293 C>T) found within intron 11 had a strong (P < 0.0005), but not complete, association with LP and CSNB and thus is a good marker but unlikely to be causative. To further localize the association, 70 SNPs spanning over two Mb including the TRPM1 gene were genotyped in 192 horses from three different breeds segregating for LP. A single 173 kb haplotype associated with LP and CSNB (ECA1: 108,197,355- 108,370,150) was identified. Illumina sequencing of 300 kb surrounding this haplotype revealed 57 SNP variants. Based on their localization within expressed sequences or regions of high sequence conservation across mammals, six of these SNPs were considered to be the most likely candidate mutations. While the precise function of TRPM1 remains to be elucidated, this work solidifies its functional role in both pigmentation and night vision. Further, this work has identified several potential regulatory elements of the TRPM1 gene that should be investigated further in this and other species.
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| 2. |
- Buijs, J, et al.
(författare)
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SPR-MS in Functional Proteomics
- 2005
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Ingår i: Briefings in Functional Genomics and Proteomics. - : Oxford University Press (OUP). - 1473-9550 .- 1477-4062. ; 4:1, s. 39-47
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Tidskriftsartikel (refereegranskat)abstract
- The mapping of protein networks and the establishment of the functional relationships between expressed proteins and their effects on cellular processes represents a great challenge for functional or interaction proteomics. The combination of surface plasmon resonance (SPR)-based technology with mass spectrometry (MS) has created a unique analytical tool for functional proteomics investigations. Proteins are affinity purified, quantified and characterised in terms of their interactions, while the mass spectrometer identifies and structurally characterises the biomolecules. Recent developments have led to a closer integration of these key technologies, providing a combined approach which enables identification of proteins selected on the basis of their functional binding criteria. In addition to a historical overview of this field, some recent detailed examples of combined SPR-MS approaches will be reviewed in a number of key application areas, including ligand fishing, peptide sequence and post-translational modification analysis by SPR-MS/MS and enzyme inhibitor screening.
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