为分析复合益生菌饲喂肉鸡回肠转录组中的新转录本预测、可变剪接事件和SNP,选择200只1日龄黄麻肉鸡随机分为2组,每组5个重复,每个重复20只.对照组正常饮水,益生菌组饮用水中补充1%复合菌制剂,试验分为生长前期(1~21 d)、后期(22~42 d)两个阶段.42日龄时进行肉鸡屠宰试验,取对照组和益生菌组回肠各3个样本进行转录组测序.对测序数据进行分析,共发现1047个新基因,20176个新转录本,276个新基因在GO数据库中得到归类注释.对转录组数据进行可变剪接分析,外显子跳跃(SE)占可变剪接类型的比例最高,共筛选到1131个显著的差异剪接基因,在外显子跳跃(SE)、第一个外显子可变剪接(A5SS)、最后一个外显子可变剪接(A3SS)、外显子选择性跳跃(MXE)、内含子滞留(RI)事件中鉴定到的差异剪接基因数分别为588、139、176、136、92.对获得的差异剪接基因进行GO富集分析,主要富集在代谢和免疫GO term.KEGG富集到了胞吞作用、MAPK信号通路等.在各样品中大多数碱基取代均为转换大于颠换,其中转换单核苷酸多态性(SNP)所占百分比为73.53%~73.94%;颠换型SNP所占百分比为26.05%~26.47%.试验对对照组、益生菌组肉鸡的回肠进行了RNA-seq测序分析,为发掘复合益生菌作用于肉鸡回肠中的可变剪接事件提供基础,为进一步了解复合益生菌作用于肉鸡回肠中新转录本的发现和SNP位点提供了数据支撑.
The purpose of the experiment was to analyze the prediction of new transcripts,alternative splicing events and SNPs in the ileal transcriptome of compound probiotic-fed broilers.200 1-day-old jute broilers were randomly divided into 2 groups with 5 replicates in each group and 20 in each replicate.The control group had normal drinking water,and the probiotic group's drinking water was supplemented with a 1%compound bacteria preparation.The test was divided into two stages:the early growth period(1~21 days)and the late growth period(22~42 days).At 42 days of age,a broiler slaughter test was carried out.Three samples from each of the ileum of the control group and the probiotic group were taken for transcriptome sequencing.The sequencing data were analyzed,a total of 1047 new genes,20176 new transcripts were found,and 276 new genes were classified and annotated in the GO database.Performing alternative splicing analysis on transcriptome data,exon skipping(SE)accounts for the highest proportion of alternative splicing types.The results of 1131 significant differences were screened.Spliced genes,Skipped exon(SE),Alternative 5'splice site(A5SS),Alternative 3'splice site(A3SS),Mutually exclusion exon(MXE),Retained intron(RI)event were 588,139,176,136,and 92,respectively.GO enrichment analysis was performed on the obtained differentially Spliced genes,mainly enriched in metabolic and immune GO terms.KEGG was enriched to endocytosis,MAPK signaling pathway and so on.In each sample,most of the base substitutions were conversion greater than transversion,and the percentage of converted SNPs was 73.53%~73.94%;the percentage of transversion SNPs was 26.05%~26.47%.RNA-seq sequencing analysis was performed on the ileum of broilers in the control group and the probiotic group,which provided a basis for exploring the alternative splicing events of the ileum of broilers acted by compound probiotics,and provided data support for further understanding the discovery of new transcripts and SNP sites of the ileum of broilers acted by compound probiotics.