小反刍兽疫病毒(PPRV)F基因在昆虫细胞中的分泌表达及免疫原性研究

doi:10.16431/j.cnki.1671-7236.2017.11.031

Abstract

Abstract:

This study was aimed to explore the use of baculovirus expression system to secrete peste des petits ruminants virus (PPRV) F gene protein and use it as subunit vaccine. The gene fragment encoding F protein of PPRV was cloned into the baculovirus pFastBac Ⅰ transfer vector with a honeybee melittin signal peptide.The constructed F-pFastBac was transformed into Escherichia coli DH10Bac,resulting the recombinant baculovirus DNA (F-Bacmid) which was confirmed by blue-white plaque assay and antibiotic resistance selection.The F-Bacmid was then transfected into Sf9 insect cells by the cellfectin transfection reagent.The recombinant F protein was expressed in High Five cells in the serum-free medium.The SDS-PAGE and Western blotting analysis of recombinant protein showed that the protein could be expressed in insect cells and secreted into the culture medium. For the immunogenicity study,the recombinant protein was then inoculated into BALB/c mice, the results showed that the recombinant protein was able to stimulate B cells to produce special antibodies. In conclusions,the recombinant baculovirus expressing F protein of PPRV were successfully constructed.This study applied a basis for the development of PPRV subunit vaccine.

Key words: PPRV F protein; baculovirs; secretory expression; immunogenicity; protective efficacy

CLC Number:

S852.65

XUE Zhong, DONG Shu-hong, WANG Shan-hui. Study on Secretory Expression and Immunogenicity of PPRV F Gene in Insect Cells[J]. , 2017, 44(11): 3327-3333.

References

[1] 世界动物卫生组织. 国际动物卫生法典[M]. 北京:中国农业科学技术出版社, 2002.
[2] 高华峰,赵文华,高林,等. 小反刍兽疫病毒原核表达体外转录系统的建立[J]. 动物医学进展,2016, 37(10):21-25.
[3] 吴锦艳,尚佑军,田宏,等. 2007-2014年国内外小反刍兽疫流行现状及分析[J]. 中国兽医学报, 2016, 36(4):687-693.
[4] 金红岩,隋修锟,刘欢,等. 抗小反刍兽疫病毒P蛋白单链抗体的制备及鉴定[J]. 中国兽医科学, 2016, 46(3):1226-1232.
[5] 邢超,苏文,王玉田,等. 一起羊小反刍兽疫的诊断及其病毒分离、鉴定和全基因组序列分析[J]. 中国兽医学报,2016, 36(10):1686-1691.
[6] Kathy H,S omeet N, Sowmya P, et al. False positive reactivity of recombinant, diagnostic, glycoproteins produced in High Five insect cell:Effect of glycosylation[J]. Immunol Methods, 2008, 330(1-2):130-136.
[7] 闫飞虎,冯娜,王琪,等. 小反刍兽疫病毒M蛋白的原核表达及多克隆抗体的制备[J]. 中国兽医学报,2016, 36(10):412-418.
[8] 李翠翠,孙一瑞,王子龙,等. 稳定表达小反刍兽疫病毒N蛋白的CHO细胞系的建立及表达产物的抗原性鉴定[J].中国预防兽医学报,2011, 33(4):262-265.
[9] 李继东,蒙学莲,朱学亮,等. 小反刍兽疫病毒H基因或F基因重组山羊痘病毒活载体疫苗的研究[J]. 中国兽医科学,2015, 45(2):111-117.
[10] 刘文华,王志亮,吴晓东,等. 小反刍兽疫病毒化学合成表位多肽对小鼠的免疫效果研究[J]. 畜牧兽医学报,2015, 46(3):438-444.
[11] van der Geld Y M, Smook M L, Huitema M G, et al. Expression of recombinant proteinase 3 the autoantigen in Wegene?s granulomatosis, in insect cells[J]. J Immunol Methods, 2002, 264(1/2):195-205.
[12] 王净,李刚,史利军. 间接ELISA和竞争ELISA检测小反刍兽疫病毒抗体的比较[J]. 畜牧兽医学报,2014, 45(3):333-336.
[13] 马学虎,马旭升,付钰广,等. 小反刍兽疫病毒China/Tib/07株V蛋白的原核表达及纯化[J]. 中国兽医科学,2013, 43(11):1140-1145.
[14] 黄华欣,李刚,史利军,等.小反刍兽疫病毒M蛋白主要抗原表位区的原核表达及鉴定[J].中国兽医科学,2013,43(3):266-270.
[15] 高华峰,信爱国,高林,等. 小反刍兽疫病毒H基因在杆状病毒中的表达[J]. 中国动物传染病学报,2012, 20(1):43-46.
[16] Futatsumori-Sugai M,Tsumoto K. Signal peptide for improving recombinant protein secretion in the baculovirus expression vector system[J]. Biochem, 2010, 391(1):931-935.
[17] Dlallo A. Control of peste des petits ruminats:Classical and new generation vaccines[J]. Dev Biol(Basel), 2003, 114:113-119.
[18] Berhe G,Minet C,Le Goff C,et al. Development of a dual recombinant vaccine to protect small ruminants against peste des petits ruminants virus and capripoxvirus infections[J]. J Virol, 2003, 77(2):1571-1577.
[19] 杨香芳,窦永喜,王秋霞,等. 小反刍兽疫病毒F基因核酸疫苗的构建及免疫原性[J]. 中国兽医科学,2013, 43(5):500-504.
[20] Monique M,Oers V. Opportunities and challenges for the baculovirus expression system[J]. Journal of Invertebrate Pathology, 2011, 107:S3-S15.

Study on Secretory Expression and Immunogenicity of PPRV F Gene in Insect Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Zhewei, DAI Xiaotong, WEI Zudan, CHEN Yunpeng, GAN Jing, BEI Weicheng. Study on Prokaryotic Expression and Immunogenicity of OmpW and TbpA Proteins in Swine Pasteurella multocida [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1489-1498.
[2]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[3]	JIANG Xiaoming, TANG Cheng, YUE Hua. Expression and Immunogenicity of P[1] Bovine Rotavirus A VP8 Recombinant Protein [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1970-1976.
[4]	LIU Qingqing, WU Peng, LI Peidong, ZHANG Jiangwei, CHEN Chuangfu, XIAO Chencheng. Construction of Recombinant Adenovirus Expressing Porcine Reproductive and Respiratory Syndrome Virus N Protein and Its Immunogenicity Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1024-1031.
[5]	ZHANG Yanmin, ZHOU Yanan, CAO Lei, TIAN Yuan, LIU Xuping, TAN Wensong, ZHAO Liang. Expression and Immunogenicity Analysis of Classical Swine Fever Virus E2-GM-CSF Fusion Protein in HEK293T Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 669-676.
[6]	ZHAO Jing, CHU Ying, LUO Jun, GUO Xiaofeng. Construction and Immunogenicity of Recombinant Rabies Virus with Canine Distemper Virus H Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4776-4785.
[7]	WU Tongzhong, NUERSAILIKE Nusufu, HUANG Xin, HAN Mengli, ZHANG Xingxing, ZHANG Qian, WANG Xinhua, HE Yanhua, ZHONG Fagang. Construction of Recombinant Lactococcus Expressing E2 Protein of Bovine Viral Diarrhea Virus and Immunogenicity Analysis of the Protein [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2975-2981.
[8]	CUI Yi, WANG Zhenhua, ZHAO Sujuan, XU Jinpeng, YANG Kai, MA Xingshu. Construction and Biological Characteristics of aro A Gene Deletion Strain of Salmonella Enteritis from Poultry [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2247-2254.
[9]	DU Jige, YAO Wensheng, LIU Ying, XUE Qi, LI Qianlin, ZHU Zhen, LI Qihong, YIN Chunsheng, CHEN Xiaoyun, LI Xuni. Expression and Immunogenicity of Recombinant Clostridium botulinum Neurotoxin Type C [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1381-1387.
[10]	LIU Zengsu, WANG Zhaorui, WANG Xueying, JIANG Yanping, CUI Wen, WANG Li, QIAO Xinyuan, ZHOU Han, XU Yigang, LI Yijing, TANG Lijie. Secretory Expression and Antiviral Activity Analysis of Recombinant Bovine Interferon-α [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1449-1456.
[11]	MENG Weijin, FAN Kuohai, CHANG Xiongxiong, YIN Wei, LI Hongquan, WANG Zhirui. Eukaryotic Expression and Immunogenicity Detection of Fusion Protein of Chicken Infectious Bursal Disease Virus VP2 and Truncated Diphtheria Toxin DT390 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4652-4660.
[12]	YANG Huihu, HUANG Huilan, YUAN Sheng, LI Wenjun, YAO Xinyan, ZHANG Yuqian, LIANG Zhaoping, HUANG Shujian, ZHANG Xuelian. Prokaryotic Expression and Immunogenicity Analysis of the Structural Protein σB of a Novel Duck Reovirus DH13 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4204-4212.
[13]	CHENG Yi, TIAN Hui, ZHANG Ya, XUE Jingjing, SHENG Dongbei, LIU Wujie, WANG Wanbing, ZHANG Pantao, TIAN Kegong. Isolation,Identification and Immunogenicity Evaluation of H9N2 Subtype Avian Influenza Virus HF Strain [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2945-2952.
[14]	ZHANG Mingliang, SUN Changjiang, GU Jingmin, CUI Ziyin, HAN Wenyu. Immunogenicity of Recombinant Salmonella Typhimurium Expressing Protective Antigen Gene F1I2 of Pseudomonas aeruginosa in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1156-1162.
[15]	HUANG Xuebing, YANG Peijie, DENG Rusen, YANG Bo, LIU Wenjun, HUANG Yunmao. Preparation of P-partical-GnIH Antigen of Norovirus and Preliminary Detection of Its Antigenicity [J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2699-2706.