PolyI:C刺激豚鹿(Axis porcinus)外周血淋巴细胞转录组分析

doi:10.16036/j.issn.1000-2650.2021.01.017

摘要/Abstract

摘要： 【目的】初步探究豚鹿的抗病毒免疫机制,为野生动物保护提供新的思路。【方法】利用转录组测序技术对聚肌胞苷酸(PolyI:C)刺激后的豚鹿外周血淋巴细胞进行测序分析并使用qPCR方法对数据的准确性进行验证。【结果】拼接得到12 1204条unigene,平均长度1 121 bp。共有97.12%的unigene得到成功注释。经过差异表达分析,获得402个差异表达基因(DEGs)。GO和KEGG富集分析显示差异基因在免疫反应方面出现富集,进而筛选出28个免疫相关基因。随机选取9个基因进行实时荧光定量分析,验证结果与测序结果变化趋势一致。【结论】获得了Poly I:C刺激后豚鹿外周血淋巴细胞的基因表达谱,为豚鹿抗病毒免疫应答机制的研究奠定了基础,为疫苗佐剂的开发及应用提供了理论基础。

关键词: Poly I:C, 豚鹿, 外周血淋巴细胞(PBL), 转录组, qPCR

Abstract: 【Objective】 The purpose of this study is to initially explore the antiviral immune mechanism of hog deer and provide new ideas for wildlife protection.【Method】 RNA-seq was used to analyze the peripheral blood lymphocytes of hog deer after poly I:C stimulation and the accuracy of the data was verified by qPCR.【Result】 121204 unigenes were spliced,with an average length was 1121 bp.After functional annotations,97.12% unigenes were successfully annotated.Through DEG-seq analysis,402 DEGs were obtained.The DEGs were subjected to enrichment analysis of GO and KEGG,and then 28 immune-related genes were screened out.9 immune-related genes were selected for qPCR verification,and the results were consistent with the trend of sequencing.【Conclusion】 This study obtained the gene expression profile after Poly I:C stimulation,which laid the basic data for the study of the immune response mechanism in the antiviral defense of hog deer,and provided a theoretical basis for the development and application of vaccine adjuvants.

Key words: PolyI:C, hog deer, peripheral blood lymphocytes (PBL), transcriptome, qPCR

中图分类号:

Q952

隋维恺, 高艳, 杨茜茜, 冷思竹, 严慧娟, 屈羽, 陈维刚, 杜小刚. PolyI:C刺激豚鹿(Axis porcinus)外周血淋巴细胞转录组分析[J]. 四川农业大学学报, 2021, 39(1): 114-120.

SUI Weikai, GAO Yanfeng, YANG Xixi, LENG Sizhu, YAN Huijuan, QU Yu, CHEN Weigang, DU Xiaogang. Transcriptome Analysis of Hog Deer (Axis Porcinus) Peripheral Blood Lymphocyte after PolyI:C Challenge[J]. Journal of Sichuan Agricultural University, 2021, 39(1): 114-120.

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