Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinärstr ,13, 80539 Munich, Germany.
Genet Sel Evol. 2011 Feb 24;43(1):8. doi: 10.1186/1297-9686-43-8.
In a previous study in the Fleckvieh dual purpose cattle breed, we mapped a quantitative trait locus (QTL) affecting milk yield (MY1), milk protein yield (PY1) and milk fat yield (FY1) during first lactation to the distal part of bovine chromosome 5 (BTA5), but the confidence interval was too large for positional cloning of the causal gene. Our objective here was to refine the position of this QTL and to define the candidate region for high-throughput sequencing.
In addition to those previously studied, new Fleckvieh families were genotyped, in order to increase the number of recombination events. Twelve new microsatellites and 240 SNP markers covering the most likely QTL region on BTA5 were analysed. Based on haplotype analysis performed in this complex pedigree, families segregating for the low frequency allele of this QTL (minor allele) were selected. Single- and multiple-QTL analyses using combined linkage and linkage disequilibrium methods were performed.
Single nucleotide polymorphism haplotype analyses on representative family sires and their ancestors revealed that the haplotype carrying the minor QTL allele is rare and most probably originates from a unique ancestor in the mapping population. Analyses of different subsets of families, created according to the results of haplotype analysis and availability of SNP and microsatellite data, refined the previously detected QTL affecting MY1 and PY1 to a region ranging from 117.962 Mb to 119.018 Mb (1.056 Mb) on BTA5. However, the possibility of a second QTL affecting only PY1 at 122.115 Mb was not ruled out.
This study demonstrates that targeting families segregating for a less frequent QTL allele is a useful method. It improves the mapping resolution of the QTL, which is due to the division of the mapping population based on the results of the haplotype analysis and to the increased frequency of the minor allele in the families. Consequently, we succeeded in refining the region containing the previously detected QTL to 1 Mb on BTA5. This candidate region contains 27 genes with unknown or partially known function(s) and is small enough for high-throughput sequencing, which will allow future detailed analyses of candidate genes.
在之前对弗莱维赫乳肉兼用牛品种的研究中,我们将影响初乳产奶量(MY1)、乳蛋白产量(PY1)和乳脂产量(FY1)的数量性状基因座(QTL)映射到牛 5 号染色体(BTA5)的远端,但该置信区间对于定位该基因的因果基因太大。我们的目标是细化该 QTL 的位置,并确定高通量测序的候选区域。
除了之前研究过的,还对新的弗莱维赫家系进行了基因分型,以增加重组事件的数量。分析了 12 个新的微卫星和 240 个覆盖 BTA5 上最有可能的 QTL 区域的 SNP 标记。基于在这个复杂家系中进行的单倍型分析,选择了该 QTL 低频等位基因(次要等位基因)分离的家系。使用连锁和连锁不平衡联合的方法进行单和多 QTL 分析。
对代表家系的父系及其祖先的单核苷酸多态性单倍型分析表明,携带次要 QTL 等位基因的单倍型很少,很可能来源于图谱群体中的一个独特祖先。根据单倍型分析的结果和 SNP 和微卫星数据的可用性,对不同的家系子集进行分析,将之前检测到的影响 MY1 和 PY1 的 QTL 细化到 BTA5 上的 117.962 Mb 到 119.018 Mb(1.056 Mb)的区域。然而,不能排除在 122.115 Mb 处仅影响 PY1 的第二个 QTL 的可能性。
本研究表明,针对分离较少的 QTL 等位基因的家系是一种有用的方法。它提高了 QTL 的作图分辨率,这是由于基于单倍型分析的结果对作图群体进行了划分,以及家系中次要等位基因的频率增加。因此,我们成功地将之前检测到的 QTL 细化到 BTA5 上的 1 Mb 区域。该候选区域包含 27 个具有未知或部分已知功能的基因,且足够小,可进行高通量测序,这将允许对候选基因进行未来的详细分析。