Qi Haigang, Song Kai, Li Chunyan, Wang Wei, Li Busu, Li Li, Zhang Guofan
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.
PLoS One. 2017 Mar 22;12(3):e0174007. doi: 10.1371/journal.pone.0174007. eCollection 2017.
Single nucleotide polymorphisms (SNPs) are widely used in genetics and genomics research. The Pacific oyster (Crassostrea gigas) is an economically and ecologically important marine bivalve, and it possesses one of the highest levels of genomic DNA variation among animal species. Pacific oyster SNPs have been extensively investigated; however, the mechanisms by which these SNPs may be used in a high-throughput, transferable, and economical manner remain to be elucidated. Here, we constructed an oyster 190K SNP array using Affymetrix Axiom genotyping technology. We designed 190,420 SNPs on the chip; these SNPs were selected from 54 million SNPs identified through re-sequencing of 472 Pacific oysters collected in China, Japan, Korea, and Canada. Our genotyping results indicated that 133,984 (70.4%) SNPs were polymorphic and successfully converted on the chip. The SNPs were distributed evenly throughout the oyster genome, located in 3,595 scaffolds with a length of ~509.4 million; the average interval spacing was 4,210 bp. In addition, 111,158 SNPs were distributed in 21,050 coding genes, with an average of 5.3 SNPs per gene. In comparison with genotypes obtained through re-sequencing, ~69% of the converted SNPs had a concordance rate of >0.971; the mean concordance rate was 0.966. Evaluation based on genotypes of full-sib family individuals revealed that the average genotyping accuracy rate was 0.975. Carrying 133 K polymorphic SNPs, our oyster 190K SNP array is the first commercially available high-density SNP chip for mollusks, with the highest throughput. It represents a valuable tool for oyster genome-wide association studies, fine linkage mapping, and population genetics.
单核苷酸多态性(SNPs)在遗传学和基因组学研究中被广泛应用。太平洋牡蛎(Crassostrea gigas)是一种在经济和生态方面都很重要的海洋双壳贝类,它在动物物种中拥有最高水平的基因组DNA变异。太平洋牡蛎的SNPs已被广泛研究;然而,这些SNPs以高通量、可转移且经济的方式应用的机制仍有待阐明。在此,我们使用Affymetrix Axiom基因分型技术构建了一个牡蛎190K SNP芯片。我们在芯片上设计了190,420个SNPs;这些SNPs是从通过对在中国、日本、韩国和加拿大收集的472个太平洋牡蛎进行重测序鉴定出的5400万个SNPs中挑选出来的。我们的基因分型结果表明,133,984个(70.4%)SNPs是多态性的并且在芯片上成功转化。这些SNPs均匀分布在整个牡蛎基因组中,位于3595个支架上,长度约为5.094亿;平均间隔距离为4210 bp。此外,111,158个SNPs分布在21,050个编码基因中,每个基因平均有5.3个SNPs。与通过重测序获得的基因型相比,约69%的转化SNPs的一致性率>0.971;平均一致性率为0.966。基于全同胞家系个体基因型的评估显示,平均基因分型准确率为0.975。我们的牡蛎190K SNP芯片携带133K个多态性SNPs,是首个可商业获得的用于软体动物的高密度SNP芯片,通量最高。它是牡蛎全基因组关联研究、精细连锁图谱绘制和群体遗传学的宝贵工具。