The Fish Molecular Genetics and Biotechnology Laboratory, Department of Fisheries and Allied Aquacultures and Program of Cell and Molecular Biosciences, Aquatic Genomics Unit, Auburn University, 203 Swingle Hall, Auburn, AL 36849, USA.
BMC Genomics. 2013 Dec 30;14:929. doi: 10.1186/1471-2164-14-929.
The application of RNA-seq has accelerated gene expression profiling and identification of gene-associated SNPs in many species. However, the integrated studies of gene expression along with SNP mapping have been lacking. Coupling of RNA-seq with bulked segregant analysis (BSA) should allow correlation of expression patterns and associated SNPs with the phenotypes.
In this study, we demonstrated the use of bulked segregant RNA-seq (BSR-Seq) for the analysis of differentially expressed genes and associated SNPs with disease resistance against enteric septicemia of catfish (ESC). A total of 1,255 differentially expressed genes were found between resistant and susceptible fish. In addition, 56,419 SNPs residing on 4,304 unique genes were identified as significant SNPs between susceptible and resistant fish. Detailed analysis of these significant SNPs allowed differentiation of significant SNPs caused by genetic segregation and those caused by allele-specific expression. Mapping of the significant SNPs, along with analysis of differentially expressed genes, allowed identification of candidate genes underlining disease resistance against ESC disease.
This study demonstrated the use of BSR-Seq for the identification of genes involved in disease resistance against ESC through expression profiling and mapping of significantly associated SNPs. BSR-Seq is applicable to analysis of genes underlining various performance and production traits without significant investment in the development of large genotyping platforms such as SNP arrays.
RNA-seq 的应用加速了许多物种的基因表达谱分析和与基因相关的 SNP 鉴定。然而,基因表达与 SNP 图谱的综合研究还很缺乏。RNA-seq 与 bulked segregant analysis(BSA)的结合应该能够将表达模式和相关 SNP 与表型相关联。
在这项研究中,我们展示了 bulked segregant RNA-seq(BSR-Seq)在分析鱼类肠败血症(ESC)抗性相关差异表达基因和 SNP 方面的应用。在抗性和易感鱼类之间发现了 1255 个差异表达基因。此外,在易感和抗性鱼类之间,鉴定出了 56419 个位于 4304 个独特基因上的 SNP 作为显著 SNP。对这些显著 SNP 的详细分析允许区分由遗传分离引起的显著 SNP 和由等位基因特异性表达引起的显著 SNP。显著 SNP 的映射,以及差异表达基因的分析,确定了 ESC 疾病抗性相关候选基因。
这项研究通过表达谱分析和显著相关 SNP 的图谱绘制,展示了 BSR-Seq 在鉴定 ESC 抗性相关基因方面的应用。BSR-Seq 适用于分析各种性能和生产性状的基因,而无需在 SNP 阵列等大型基因分型平台的开发方面进行大量投资。