猫importin基因生物信息分析及其在犬细小病毒感染F81细胞表达的动态变化

doi:10.16431/j.cnki.1671-7236.2020.12.008

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (12): 3861-3869.doi: 10.16431/j.cnki.1671-7236.2020.12.008

猫importin基因生物信息分析及其在犬细小病毒感染F81细胞表达的动态变化

赵航¹, 宋姗姗², 王建科³, 林鹏⁴

1. 江苏省农业科学院农产品质量安全与营养研究所, 江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地, 南京 210014;
2. 中国农业科学院特产研究所, 吉林省特种经济动物分子生物学省部共建国家重点实验室, 长春 130112;
3. 中国农业科学院特产研究所, 特种经济动物分子生物学国家重点实验室, 长春 130112;
4. 渤海大学食品科学与工程学院, 生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心, 国家鱼糜及鱼糜制品加工技术研发分中心, 锦州 121013

收稿日期:2020-07-06 出版日期:2020-12-20 发布日期:2020-12-18
通讯作者: 林鹏 E-mail:lphs1002@163.com
作者简介:赵航(1988-),女,黑龙江哈尔滨人,博士,助理研究员,研究方向:病毒致病机制,E-mail:dongyizhaohang@163.com
基金资助:
国家自然科学基金（3170131433）

Bioinformatics Analysis of Feline importin Gene and Its Dynamic Expression in F81 Cells Infected with Canine Parvovirus

ZHAO Hang¹, SONG Shanshan², WANG Jianke³, LIN Peng⁴

1. Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base, Key Laboratory of Agro-food Safety and Quality, Institute of Food Safety and Detection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. Jilin Province State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;
3. State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;
4. National & Local Joint Engineering Research Center of Storage, Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products, National R & D Branch Center of Surimi and Surimi Products Processing, College of Food Science and Technology, Bohai University, Jinzhou 121013, China

Received:2020-07-06 Online:2020-12-20 Published:2020-12-18

摘要/Abstract

摘要： 试验旨在系统研究猫源不同亚型importin（importin α1、importin α3、importin α4、importin α5、importin α6、importin α7、importin α8和importin β）基因序列和蛋白基本特性，探讨其在犬细小病毒（canine parvovirus，CPV）感染F81细胞表达的动态变化。本研究利用RT-PCR方法扩增F81细胞中improtin基因全长序列，应用生物学软件预测其氨基酸序列及编码蛋白的结构及功能；进一步利用实时荧光定量RT-PCR技术检测importin基因在CPV感染F81细胞后24、48和72 h表达水平。结果显示，不同亚型importin α基因全长约在1 500 bp，而importin β基因全长为2 600 bp；预测importin亚型蛋白序列发现，其等电点均在4.60左右，且发现importin α1、importin α5、importin α6、importin α7和importin β为不稳定蛋白；分析发现这些importin蛋白均无信号肽、无跨膜区、无细胞定位；采用Predictprotein软件预测蛋白二级结构，结果显示importin亚型蛋白序列主要以α-螺旋和无规则卷曲形式存在。实时荧光定量RT-PCR结果显示，CPV感染F81细胞后随着感染时间的延长importin α1（P<0.01）和importin β表达量逐渐降低，而importin α3、importin α4、importin α5、importin α6（P<0.01）、importin α7（P<0.01）和importin α8（P<0.01）表达量逐渐提高，其中importin α8在病毒感染细胞后表达量最高。本研究结果为进一步分析CPV进入细胞核的转运机制及开发治疗药物相关研究奠定基础。

关键词: 输入蛋白; 犬细小病毒(CPV); 序列分析; 表达差异

Abstract: This study was aimed to systematically analyze the gene sequences and proteins of the importin subtype genes (importin α1,importin α3,importin α4,importin α5,importin α6,importin α7,importin α8 and importin β),and the change of transcription level of importin subtype of F81 cells infected CPV.The full length of importin genes were amplified by RT-PCR techniques from F81 cells and the amino acid sequence and the structure and function of importin protein were subsequently analyzed by bioinformatics software.Meanwhile,the dynamic change of expression of importin genes in CPV-infected F81 cells were investigated at 24,48 and 72 h postinfection by Real-time RT-PCR.The results showed that full length of the importin α subtype genes were about 1 500 bp and the importin β was 2 600 bp.The physical chemical properties of importin proteins showed the theoretical isoelectric was about 4.60 and the importin α1,importin α5,importin α6,importin α7 and importin β belonged to instable proteins.Meanwhile,there were no signal peptide,no transmembrane structures and no cellular localization in importin proteins.The secondary structure of importin proteins were predicted by Predictprotein software and mainly composed of α-helix and random coil.Real-time RT-PCR results showed that the importin α1 (P<0.01) and importin β mRNA level were significantly reduced in CPV-infected F81 cells,while other importin mRNA level were markedly up-increased,the importin α8 gene had the highest expressional level (P<0.01).The study laid a foundation for further study on the transport mechanism of CPV into the nucleus of the cell and developing drugs.

Key words: importin; canine parvovirus (CPV); sequence analysis; expression difference

中图分类号:

829.3

赵航, 宋姗姗, 王建科, 林鹏. 猫importin基因生物信息分析及其在犬细小病毒感染F81细胞表达的动态变化[J]. 中国畜牧兽医, 2020, 47(12): 3861-3869.

ZHAO Hang, SONG Shanshan, WANG Jianke, LIN Peng. Bioinformatics Analysis of Feline importin Gene and Its Dynamic Expression in F81 Cells Infected with Canine Parvovirus[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 3861-3869.

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猫importin基因生物信息分析及其在犬细小病毒感染F81细胞表达的动态变化

Bioinformatics Analysis of Feline importin Gene and Its Dynamic Expression in F81 Cells Infected with Canine Parvovirus

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