Departament de Ciència Animal i dels Aliments, Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Heredity (Edinb). 2011 Sep;107(3):256-64. doi: 10.1038/hdy.2011.13. Epub 2011 Mar 16.
Despite dramatic reduction in sequencing costs with the advent of next generation sequencing technologies, obtaining a complete mammalian genome sequence at sufficient depth is still costly. An alternative is partial sequencing. Here, we have sequenced a reduced representation library of an Iberian sow from the Guadyerbas strain, a highly inbred strain that has been used in numerous QTL studies because of its extreme phenotypic characteristics. Using the Illumina Genome Analyzer II (San Diego, CA, USA), we resequenced ∼ 1% of the genome with average 4 × depth, identifying 68,778 polymorphisms. Of these, 55,457 were putative fixed differences with respect to the assembly, based on the genome of a Duroc pig, and 13,321 were heterozygous positions within Guadyerbas. Despite being highly inbred, the estimate of heterozygosity within Guadyerbas was ∼ 0.78 kb(-1) in autosomes, after correcting for low depth. Nucleotide variability was consistently higher at the telomeric regions than on the rest of the chromosome, likely a result of increased recombination rates. Further, variability was 50% lower in the X-chromosome than in autosomes, which may be explained by a recent bottleneck or by selection. We divided the whole genome in 500 kb windows and we analyzed overrepresented gene ontology terms in regions of low and high variability. Multi organism process, pigmentation and cell killing were overrepresented in high variability regions and metabolic process ontology, within low variability regions. Further, a genome wide Hudson-Kreitman-Aguadé test was carried out per window; overall, variability was in agreement with neutral expectations.
尽管随着新一代测序技术的出现,测序成本大幅降低,但要获得足够深度的完整哺乳动物基因组序列仍然很昂贵。另一种选择是部分测序。在这里,我们对来自 Guadyerbas 品系的伊比利亚母猪的简化代表文库进行了测序,Guadyerbas 品系是一种高度近交的品系,由于其极端的表型特征,已被用于许多 QTL 研究中。使用 Illumina Genome Analyzer II(加利福尼亚州圣地亚哥),我们以平均 4×的深度重新测序了基因组的约 1%,鉴定出 68778 个多态性。其中,根据杜洛克猪的基因组,有 55457 个是相对于组装的假定固定差异,有 13321 个是 Guadyerbas 中的杂合位置。尽管高度近交,但在 Guadyerbas 中,经过对低深度的校正,每个个体的杂合度估计约为 0.78 kb(-1)。端粒区域的核苷酸变异性始终高于染色体的其余部分,这可能是由于重组率增加所致。此外,X 染色体的变异性比常染色体低 50%,这可能是由于最近的瓶颈或选择造成的。我们将整个基因组分为 500 kb 的窗口,并分析低变异性和高变异性区域中基因本体论术语的过度表达。在高变异性区域中,多器官过程、色素沉着和细胞杀伤过度表达,而在低变异性区域中,代谢过程本体论过度表达。此外,对每个窗口进行了全基因组 Hudson-Kreitman-Aguadé 测试;总体而言,变异性与中性预期一致。