基于RNA-Seq筛选鸭肠炎病毒感染鸭脾差异表达免疫相关基因

doi:10.11843/j.issn.0366-6964.2017.02.013

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (2): 297-306.doi: 10.11843/j.issn.0366-6964.2017.02.013

基于RNA-Seq筛选鸭肠炎病毒感染鸭脾差异表达免疫相关基因

吴良涛^1，2，3，郑敏^1，2，3，华敏^1，2，3，万润^1，2，3，程振涛^1，2，3，周碧君^1，2，3，文明^1，2，3*

（1.贵州大学动物科学学院，贵阳 550025；2.贵州大学动物疫病研究所，贵阳 550025；3.贵州省动物疫病与兽医公共卫生重点实验室，贵阳 550025）

收稿日期:2016-09-20 出版日期:2017-02-23 发布日期:2017-02-23
通讯作者: 文明，博士，教授，E-mail：as.mwen@gzu.edu.cn
作者简介:吴良涛(1988-)，男，侗族，贵州榕江人，硕士生，主要从事病毒学研究，E-mail：826417155@qq.com
基金资助:
国家自然科学基金项目(31260607；31560703)；贵州省优秀青年科学人才培养计划项目(黔科合人字［2013］25号)；贵州省百层次创新型人才项目(黔科合人才［2016］4009号)；贵州省科技创新人才团队建设项目(黔科合人才团队［2015］4016号)

Screening of Differentially Expressed Immune-related Genes from Duck Spleen with Duck Enteritis Virus Infection Based on RNA-Seq Technology

WU Liang-tao^1,2,3, ZHENG Min^1,2,3, HUA Min^1,2,3, WAN Run^1,2,3, CHENG Zhen-tao^1,2,3, ZHOU Bi-jun^1,2,3, WEN Ming^1,2,3*

(1.College of Animal Science, Guizhou University, Guiyang 550025, China; 2.Institute for Animal Diseases, Guizhou University, Guiyang 550025, China; 3.Key Laboratory of Animal Diseases and Veterinary Public Health in Guizhou Province, Guiyang 550025, China)

Received:2016-09-20 Online:2017-02-23 Published:2017-02-23

摘要/Abstract

摘要：

为挖掘鸭肠炎病毒(duck enteritis virus，DEV)感染鸭的差异表达免疫相关基因，本研究以DEV GZ株经腿部肌肉接种50 d鸭后，于接种后66、90和114 h采集鸭脾组织样本，提取总RNA，通过高通量RNA-seq技术测序，应用GO和KEGG数据库进行比对注释，并采用荧光定量PCR对部分差异表达基因进行验证，结果显示：DEV接种66 h时鸭脾的差异表达基因有511个，参与免疫相关生物学过程的为70个，其中上调基因46个，下调基因24个；90 h时差异表达基因有485个，参与免疫相关生物学过程的为64个，其中上调基因49个，下调基因15个；114 h时差异表达基因有531个，参与免疫相关生物学过程的为66个，其中上调基因35个，下调基因31个。GO数据库分析显示，差异表达免疫相关基因主要涉及细胞黏附、抗原加工和呈递、补体激活等免疫反应过程；KEGG数据库分析显示，差异表达免疫相关基因主要富集在细胞黏附分子、ECM受体互作、PPAR等信号通路；荧光定量PCR对7个差异表达基因进行检测显示，差异表达基因相对表达量与RNA-Seq技术测序结果基本一致。这些结果为进一步阐明DEV感染机制和机体免疫应答机制奠定了基础。

Abstract:

The aim of the present study was to screen the differentially expressed immune-related genes from duck spleen by DEV infection. Ducks of 50 days old were selected to inoculate with DEV in leg muscle, and the duck spleens were collected at 66, 90 and 114 h after inoculation, and were extracted for total RNA, then sequencing by high-throughput RNA-Seq technology. The sequenced segments were compared and noted for screening of the differentially expressed immune-related genes by GO and KEGG database in NCBI. Some of the differentially expressed genes were selected to verify by real-time PCR technology. The results showed that there were 511 of the differentially expressed genes in duck spleen at 66 h after DEV infection, which 70 involved in the biological immune process, including 46 up-regulated genes and 24 down-regulated genes. At 90 h after DEV infection, there were 485 of the differentially expressed genes, which 64 involved in the biological immune process, including 49 up-regulated genes and 15 down-regulated genes. At 114 h after DEV infection, there were 531 of the differentially expressed genes, which 66 involved in the biological immune process, including 35 up-regulated genes and 31 down-regulated genes. The differentially expressed immune-related genes were mainly involved in cell adhesion, antigen processing and presentation and complement activation through GO database analysis. And these genes were mainly enriched in the signaling pathways as cell adhesion molecule, ECM receptor interaction and PPAR, etc. The expressed contents of the selected 7 genes by the FQ-PCR, were basically the same with the sequencing results by RNA-Seq technology. These results provided a theoretical basis for further understanding of the mechanism of DEV infection and duck immune respone.

中图分类号:

吴良涛，郑敏，华敏，万润，程振涛，周碧君，文明. 基于RNA-Seq筛选鸭肠炎病毒感染鸭脾差异表达免疫相关基因[J]. 畜牧兽医学报, 2017, 48(2): 297-306.

WU Liang-tao, ZHENG Min, HUA Min, WAN Run, CHENG Zhen-tao, ZHOU Bi-jun, WEN Ming. Screening of Differentially Expressed Immune-related Genes from Duck Spleen with Duck Enteritis Virus Infection Based on RNA-Seq Technology[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 297-306.

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基于RNA-Seq筛选鸭肠炎病毒感染鸭脾差异表达免疫相关基因

Screening of Differentially Expressed Immune-related Genes from Duck Spleen with Duck Enteritis Virus Infection Based on RNA-Seq Technology

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