近交系小鼠C57BL/6和C57BL/10的转录组差异研究

目的:对近交系小鼠C57BL/6和C57BL/10进行转录组测序,对比其转录组差异,发现免疫反应相关差异基因,为药物体内活性评价动物选择提供参考。方法:采用Illumina Hiseq 2000测序平台进行测序,通过短序列分析获取转录本,通过基因功能分类(GO)和京都基因与基因组百科全书(KEGG)数据库进行测序结果分析。结果:获得38553614个C57BL/6小鼠的原始序列和37721318个C57BL/10小鼠的原始序列。获得118002个转录本,注释了46193个差异基因。38106个转录本注释到2196个GO分类中,GO分类主要分为3类:细胞成分,分子功能和生物过程。1600个转录本注释到254个KEGG通路中。另外,发现了4651个单核苷酸(SNP)位点。结论:通过二代测序方法发现近交系小鼠C57BL/6和C57BL/10转录组存在显著差异基因。在KEGG通路分析中筛选了一系列与小鼠免疫应答相关的差异基因,为建立小鼠免疫评价模型积累了数据,对药物实验动物体内活性检测分析具有参考意义,为生物制品评价用实验动物的标准化提供技术支撑。

Objective:To sequence transcriptome profile of two inbred mouse strains C57BL/6(B6) and C57BL/10(B₁0), compare the difference in transcriptome profile and explore differential immune response related genes, in order to provide reference for animal selection for drug activity evaluation.Methods:Illumina Hiseq 2000 platform was utilized for sequencing, transcripts were acquired using de novo assembly, and differential genes were annotated by Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG).GO terms were categorized into three major functional groups:cellular component, molecular function, and biological process.Results:A total of 38553614 raw reads of B6 and 37721318 raw reads of B₁0 were obtained.118002 transcripts were acquired, and 46193 differential genes were annotated.A total of 38106 unique transcripts were assigned to 2196 GO terms.A total of 1600 transcripts were assigned to 254 KEGG pathways.Furthermore, we identified 4651 single nucleotide polymorphism(SNP) sites.Conclusions:Significantly differentially expressed genes were found between the inbred mouse strains C57BL/6(B6) and C57BL/10(B₁0), as revealed by second-generation sequencing method.High-throughput second-generation sequencing method could provide large amount of information and comprehensive analysis, and is an efficient way to study genomic differences between biological species.In KEGG pathway analysis, a series of differential genes associated with immune response in mice were identified.This study accumulated data for establishing immune response evaluation mice model, provided reference for drug activity test in animal, as well as technical support for the standardization of experimental animals in biological products evaluation.