摘要
为了探究猪瘟兔化弱毒株(Chinese strain, C株)在家兔体内的病毒清除机制,试验以新西兰兔为材料,适应性喂养7 d后随机分为3组,即1个对照组和2个试验组,每组6只。2个试验组家兔分别在感染C株后第5天和第9天耳缘静脉注射空气处死,取出脾脏,获得T1和T2样本,采用TRIzol法提取脾脏总RNA,利用mRNA-Seq技术对家兔脾脏进行转录组测序与分析,以正常家兔基因组为参考,筛选出差异表达基因并进行GO功能富集分析和KEGG信号通路分析,并利用实时荧光定量PCR方法对部分差异基因进行验证。结果表明:与对照组相比,感染C株后T1样本获得的上调基因和下调基因分别有61个和25个,T2样本分别有22个和8个。T1样本差异基因映射到109个GO条目,T2样本差异基因映射到61个GO条目,T1和T2样本差异基因均富集到代谢过程和细胞进程。T1样本差异基因主要富集在趋化因子信号通路、细胞因子与其受体通路、细胞周期通路、RIG-Ⅰ样受体信号通路和Toll样受体信号通路,T2样本差异基因主要富集在精氨酸和脯氨酸代谢通路、维生素B6代谢通路以及钙信号通路。实时荧光定量PCR验证结果与mRNA-Seq技术的分析结果基本一致。说明家兔感染C株后,T1样本主要通过免疫反应抵抗、清除病毒,T2样本主要通过细胞代谢相关基因的调控来恢复机体细胞的正常功能。
In order to explore the virus clearance mechanism of the lapinized attenuated strain of Classical swine fever virus(Chinese strain, C strain) in rabbits, New Zealand rabbits were used in this experiment as materials. After 7 days of adaptive feeding, they were randomly divided into 3 groups for C strain inoculation, namely one control group and two test groups, 6 rabbits in each group. The rabbits were sacrificed by air injection into the ear vein on the 5 th and 9 th days after C strain infection, and then the spleen was taken out, named T1 and T2 samples;and the total RNA of the spleen was extracted by TRIzol method, and the sequencing and analysis of the transcriptome of rabbit spleen were carried out by mRNA-Seq technology. Using control rabbit genome as a reference, differentially expressed genes were screened out;and GO functional enrichment analysis and KEGG signal pathway analysis were performed, and real-time fluorescent quantitative PCR method was used to verify some differential genes. The results showed that compared with the control group, the T1 samples obtained 61 up-regulated genes and 25 down-regulated genes after the infection with the C strain, respectively, and the T2 samples had 22 and 8 respectively. Differential genes in T1 samples were mapped to 109 entries, and differential genes in T2 samples were mapped to 61 entries;differential genes in T1 and T2 samples were both enriched in metabolic processes and cellular processes. Differential genes in T1 samples mainly involved chemokine signaling pathways, cytokine and their receptor pathways, cell cycle pathways, RIG-I-like signaling pathways, and Toll like receptor signaling pathways. Differential genes in T2 samples mainly involved arginine and proline Metabolic pathway, vitamin B6 metabolic pathway and renin-angiotensin system pathway. The real-time fluorescent quantitative PCR verification results were basically consistent with the analysis results of mRNA-Seq technology. The results suggested that after rabbits were infected with the C strain, T1 samples mainly resisted and cleared the virus through immune response, and T2 samples mainly restored the normal function of body cells through the regulation of cell metabolism-related genes.
作者
邓玲聪
廖青
邓吉平
纪佳雨
袁露露
许道军
DENG Lingcong;LIAO Qing;DENG Jiping;JI Jiayu;YUAN Lulu;XU Daojun(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;Institute of Biomedicine and Biotechnology,Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2021年第7期6-13,共8页
Heilongjiang Animal Science And veterinary Medicine
基金
国家自然科学基金项目(31201968)
2019年湖南省研究生科技创新项目(CX20190510)。
