Construction of Tagged Recombinant Porcine Circovirus Type 2 and Their Immunogenicity in Mice

doi:10.13304/j.nykjdb.2017.0356

Journal of Agricultural Science and Technology ›› 2018, Vol. 20 ›› Issue (4): 36-43.DOI: 10.13304/j.nykjdb.2017.0356

Construction of Tagged Recombinant Porcine Circovirus Type 2 and Their Immunogenicity in Mice

HUA Tao1,2,3, TANG Bo1,2,3, CHANG Chen1,2,3, LIU Guoyang1,2,3, ZHANG Xuehua1,2,3, YU Yang1,2,3, HOU Jibo1,2,3, ZHANG Daohua1,2,3*

1.Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base, Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014; 2.National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Sciences,Nanjing 210014; 3.Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Jiangsu Yangzhou 225009, China

Received:2017-05-31 Online:2018-04-15 Published:2017-09-01

猪圆环病毒2型标签重组病毒构建和免疫试验

华涛1,2,3,唐波1,2,3,常晨1,2,3,刘国阳1,2,3,张雪花1,2,3,于漾1,2,3,侯继波1,2,3,张道华1,2,3*

1.江苏省农业科学院动物免疫工程研究所, 省部共建国家重点实验室培育基地-江苏省食品质量安全重点实验室, 南京 210014; 2.江苏省农业科学院国家兽用生物制品工程技术研究中心, 南京 210014;3.江苏省动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009

通讯作者: 张道华，研究员，主要从事兽用生物制品工程技术和疫苗研究。E-mail：zhangdh2005@163.com
作者简介:华涛，助理研究员，博士，主要从事兽用生物制品工程技术和疫苗研究。E-mail：huatao541121@163.com。
基金资助:
江苏省农业自主创新项目\[CX(14)5044\]；农业部公益性行业（农业）科研专项经费项目(201303046)资助。

Abstract

Abstract: Porcine circovirus type 2 (PCV2) can cause postweaning multisystemic wasting syndrome. In order to obtain the purified virus and develop novel marker vaccine against PCV2, a 6-amino-acid His tag was inserted into the C-terminus of capsid protein of PCV2. The recombinant mark virus rPCV2-His expressing the His tag was constructed and rescued. The recombinant marker virus was detected by indirect immunouorescence assay and Western blot, and was purified by nickel column. PCV2 specific antibody was significantly improved and His tag immune antibody was produced in mice inmunized with purified recombinant virus. PCV2 content was significantly decreased in mice inmunized with purified recombinant virus. The findings of this study may be used for the development of a novel PCV2 vaccine.

Key words: PCV2, recombinant virus, purification, marker, immune

摘要： 猪圆环病毒2型（PCV2）能引起断奶仔猪衰竭综合症，为了获得利于纯化的病毒和新型标记疫苗，采用病毒重组技术，选择利于纯化的6个氨基酸His标签作为标记表位，插入PCV2 Cap蛋白 C端，拯救获得重组标记病毒rPCV2-His。间接免疫荧光和Western blot鉴定结果证明重组病毒所含的标签能稳定表达。通过镍柱纯化重组病毒，该病毒获得了纯化。纯化的重组病毒灭活后免疫小鼠，PCV2特异抗体显著提升，且能产生标记免疫抗体，小鼠攻毒显著降低体内PCV2含量。该重组病毒可作为PCV2新型疫苗研究重要材料。

关键词: PCV2, 重组病毒, 纯化, 标记, 免疫

HUA Tao1,2,3, TANG Bo1,2,3, CHANG Chen1,2,3, LIU Guoyang1,2,3, ZHANG Xuehua1,2,3, YU Yang1,2,3, HOU Jibo1,2,3, ZHANG Daohua1,2,3*. Construction of Tagged Recombinant Porcine Circovirus Type 2 and Their Immunogenicity in Mice[J]. Journal of Agricultural Science and Technology, 2018, 20(4): 36-43.

华涛1,2,3,唐波1,2,3,常晨1,2,3,刘国阳1,2,3,张雪花1,2,3,于漾1,2,3,侯继波1,2,3,张道华1,2,3*. 猪圆环病毒2型标签重组病毒构建和免疫试验[J]. 中国农业科技导报, 2018, 20(4): 36-43.

[1]	SUN Jing1, REN Wenqi1, XU Qinhu2, CAO Wei1, LIU Hao1. Transformation System of Aspergillus niger Using amdS as a Dominant Selective Marker [J]. Journal of Agricultural Science and Technology, 2020, 22(9): 179-187.
[2]	DUAN Min1, XIE Liujie1, ZHU Yajun2, HUANG Shanjun1, PAN Xiaobiao1, XU Jianlong2,3. salt tolerance; survival rate; SSR molecular marker; quantitative trait locus; genetic linkage mapQTL Mapping of Seedling Survival Rate Under Salt Stress in Rice (Oryza sativa L.) [J]. Journal of Agricultural Science and Technology, 2019, 21(9): 25-35.
[3]	LIU Min1, WU Tieming1*, LIU Han1, YU Mengjie1, XIAO Shasha1, HAN Qipeng2. Study on Purification of Eutrophic Water by Different Combinations of Three Aquatic Plants [J]. Journal of Agricultural Science and Technology, 2019, 21(7): 155-160.
[4]	LI Yang1,2, ZOU Junjie2, YU Jia2, XU Yu2,3, XU Miaoyun2, LUO Hongfa1, WANG Lei2. Construction of Maize Near-Isogenic Lines and Its Application [J]. Journal of Agricultural Science and Technology, 2019, 21(12): 14-22.
[5]	WU Jie1,2, GONG Jiangning3. Purification of Total Flavonoids from Flos Lonicerae by the Macroporous Resins and Its Inhibitory Activity on α-glucosidase [J]. Journal of Agricultural Science and Technology, 2019, 21(10): 89-97.
[6]	DONG Zhigang1,2, LIU Wei2, LI Xiaomei2, TAN Wei2, ZHAO Qifeng2, . Analysis of Genetic Relationship of 15 Winegrape Germplasm Resources by SSR Markers [J]. Journal of Agricultural Science and Technology, 2017, 19(9): 42-50.
[7]	OU Jiashun1,2, LIU Dawei1,2,3, LI Xu1,2,3, XIE Fangping1,2,3*, WANG Xiushan1,2, XIE Chao1,2. Design and Test of High Self-purification Threshing Device for Small Rice Combine Harvester [J]. Journal of Agricultural Science and Technology, 2017, 19(9): 63-69.
[8]	KONG Youbin1, WANG Bing1, ZHANG Hua1, LI Xihuan1, ZHANG Caiying2. Construction and Amplification of Protein Fusion HA Tag Universal Expression Vector in Plant [J]. Journal of Agricultural Science and Technology, 2017, 19(6): 131-137.
[9]	WANG Yi, HU Wan-bin, LI Jia-xiang, HUANG Xian-ya, DUAN Li-zhu, ZHAN Yao-yao*. Effects of Acute Salinity Stress on Gill structure and Four Enzyme Activities in Saxidomus purpurata [J]. Journal of Agricultural Science and Technology, 2016, 18(5): 178-186.
[10]	ZHAO Yue1,2, ZHANG Shi-lai2, HU Jian3, HUANG Li-yu4, ZHANG Jing2, . Insect-resistant Rice Cultivar Breeding by Marker-assisted Selection [J]. Journal of Agricultural Science and Technology, 2016, 18(3): 25-31.
[11]	CHEN Xiu-hua, YU Li-juan, LUO Li-ming, CHEN Hong-mei, LIU Li*. Research Progress on Maize Molecular Marker-assisted Breeding and Marker Development [J]. Journal of Agricultural Science and Technology, 2016, 18(1): 26-31.
[12]	JIANG Jing-wei, DONG Ying, ZHOU Zun-chun*. Research Status on Laccase-type Phenoloxidase in Marine Invertebrate [J]. , 2015, 17(3): 167-174.
[13]	. Identification of SCAR Molecular Marker Technology among Ganoderma lucidum Strains [J]. , 2014, 16(6): 44-51.
[14]	QU Meng-nan1,2§, JIANG Bing-jun2§, LIU Wei2, MA Li-ming2, LIN Kang-xue2, HAN T F. New Approaches to Molecular Breeding of Soybean [J]. , 2014, 16(3): 8-13.
[15]	KANG Le1, WANG Hai\\|yang2*. Status and Prospect of Agrobiotechnology\\|based Breeding in China [J]. , 2014, 16(1): 16-23.

Construction of Tagged Recombinant Porcine Circovirus Type 2 and Their Immunogenicity in Mice

猪圆环病毒2型标签重组病毒构建和免疫试验

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics