猪O型口蹄疫病毒样颗粒的构建及鉴定

doi:10.16431/j.cnki.1671-7236.2019.11.025

Abstract

Abstract: To develop the virus-like particles (VLPs) genetic engineering subunit vaccine for swine O-type foot-and-mouth disease virus (FMDV),according to the FMDV complete gene sequence available in GenBank (accession No.:JN998085),four pairs of specific primers were designed to amplify VP1,VP2,VP3 and VP4 genes of FMDV O/MYA98/XJ/2010 strain by PCR. VP3, VP1 and VP4,VP2 gene were cloned into p10 and pH promoters of pFastBac Dual vector,respectively.The recombinant plasmid pFBD-VP3-VP1 and pFBD-VP4-VP2 were obtained and identified.Two recombinant bacmid of rBacmid-VP3-VP1 and rBacmid-VP4-VP2 were generated after the E.coli DH10Bac competent cell was transformed by the two recombinant plasmid,respectively.Two recombinant baculovirus of rvAc-VP3-VP1 and rvAc-VP4-VP2 were successfully rescued by transfected two recombinant bacmids into Sf9 insect cells.Two recombinant baculoviruses were coinfected with suspension cultured Sf9 insect cells,and VP1, VP2, VP3 and VP4 genes were expressed via Bac-to-Bac expression system in Sf9 insect cells.The target proteins were identified and analyzed with specific positive antibody through IFA,SDS-PAGE and Western blotting,and the target proteins were checked as VLPs via by electron microscopy (EM).The results showed that two recombinant baculoviruses expressing four structural proteins of FMDV VP1,VP2,VP3 and VP4 were successfully constructed.The specific antibodies detection results showed that four proteins VP1,VP2,VP3 and VP4 were successfully expressed with good specificity.Four proteins could self-assemble in Sf9 insect cells and form VLPs with a diameter of 25-30 nm similar to that of natural viruses.In this study,FMDV VLPs were successfully prepared in Sf9 insect cells by coinfection expression,which laid a foundation for further developing efficient and safe FMDV gene engineering subunit vaccine.

Key words: foot-and-mouth disease virus (FMDV); structural protein; recombinant baculovirus; virus-like particles (VLPs)

CLC Number:

S852.65⁺9.6

LIU Hanping. Construction and Identification of Swine O-type Foot-and-mouth Disease Virus-like Particles[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3350-3357.

References

[1] 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997. YIN Z,LIU J H.Animal Virology[M].2nd Edition.Beijing:Science Press,1997.(in Chinese)
[2] GRUBMAN M J,BAXT B.Foot-and-mouth disease[J].Clinical Microbiology Reviews,2004,17(2):465-493.
[3] CAO Y,LU Z,LIU Z.Foot-and-mouth disease vaccines:Progress and problems[J].Expert Review of Vaccines,2016,15(6):783-789.
[4] PATON D J,KING D P,KNOWLES N J,et al.Recent spread of foot-and-mouth disease in the Far East[J].Veterinary Record,2010,166(18):569-570.
[5] DOEL T R.FMD vaccines[J]. Virus Research,2003,91(1):81-99.
[6] 张庆勋,刘新生,周鹏,等.口蹄疫病毒病毒样颗粒研究进展[J].中国畜牧兽医,2014,41(11):258-262. ZHANG Q X,LIU X S,ZHOU P,et al.Research progress on foot-and-mouth disease virus virus-like particles[J].China Animal Husbandry & Veterinary Medicine,2014,41(11):258-262.(in Chinese)
[7] 李世芳,邵军军,常惠芸.口蹄疫表位疫苗的研究进展[J].中国畜牧兽医,2017,44(2):369-401. LI S F,SHAO J J,CHANG H Y.Research progress on the epitope-based vaccine against foot and mouth disease[J].China Animal Husbandry & Veterinary Medicine,2017,44(2):369-401.(in Chinese)
[8] 李明,张雅娜,郭晓宇,等.口蹄疫疫苗非抗原蛋白对146S抗原免疫效果的影响[J].中国畜牧兽医,2019,46(3):905-912. LI M,ZHANG Y N,GUO X Y,et al.Effect of non-antigen protein of foot-and-mouth disease vaccine on immune effect of 146S antigen[J].China Animal Husbandry & Veterinary Medicine,2019,46(3):905-912.(in Chinese)
[9] TERHUJA M,SARAVANAN P,TAMILSELVAN R P.Comparative efficacy of virus like particle (VLP) vaccine of foot-and-mouth-disease virus(FMDV) type O adjuvanted with poly Ⅰ:C or CpG in guinea pigs[J].Biologicals,2015,43(6):437-443.
[10] OEM J K,PARK J H,LEE K N,et al.Characterization of recombinant foot-and-mouth disease virus pentamer-like structures expressed by baculovirus and their use as diagnostic antigens in a blocking ELISA[J].Vaccine,2007,25(20):4112-4121.
[11] 李淑萍,孙世琪,莫亚霞,等.O型口蹄疫病毒样颗粒编码基因的真核表达载体构建及表达[J].动物医学进展,2019,40(1):1-6. LI S P,SUN S Q,MO Y X,et al.Construction and expression of eukaryotic expression vector of O type foot-and-mouth disease virus-like particles encoding gene[J].Progress in Veterinary Medicine,2019,40(1):1-6.(in Chinese)
[12] CLAUDINE P,XUA X D,SILVIA L,et al.Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity[J].Journal of Virological Methods,2013,187(2):406-412.
[13] LI Z,YI Y,YIN X,et al.Expression of foot-and-mouth disease virus capsid proteins in silkworm-baculovirus expression system and its utilization as a subunit vaccine[J].PLoS One,2008,3(5):e2273.
[14] DONG H,GUO H C,SUN S Q.Virus-like particles in picornavirus vaccine development[J].Applied Microbiology and Biotechnology,2014,98(10):4321-4329.
[15] BRAUTIGAM S,SNEZHKOV E,BISHOP D H.Formation of poliovirus-like particles by recombinant baculoviruses expressing the individual VP0,VP3 and VP1 proteins by comparison to particles derived from the expressed poliovirus polyprotein[J]. Virology,1993,192(2):512-524.
[16] PUCKETTE M,CLARK B A,SMITH J D.Foot-and-mouth disease (FMD) virus 3C protease mutant L127P:Implications for FMD vaccine development[J].Journal of Virology,2017,91(22):e00924-17.
[17] MARTEL E,FORZONO E,KURKER R,et al.Effect of foot-and-mouth disease virus 3C protease B2β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector[J].Journal of General Virology,2019,100(3):446-456.
[18] 韦平,秦爱建.重要动物病毒分子生物学[M].北京:科学出版社,2008. WEI P,QIN A J.Molecular Biology of Important Animal Viruses[M].Beijing:Science Press,2008.(in Chinese)
[19] 李天芝,于新友.口蹄疫基因工程疫苗研究进展[J].猪业科学,2018,35(8):110-111. LI T Z,YU X Y.Advances in genetically engineered vaccines against foot-and-mouth disease virus[J].Advances in Swine Industry Science,2018,35(8):110-111.(in Chinese)
[20] KUSHNIR N,STREATFIELD S J,YUSIBOV V.Virus-like particles as a highly efficient vaccine platform:Diversity of targets and production systems and advances in clinical development[J].Vaccine,2012,31(1):58-83.
[21] CAO Y,SUN P,FU Y,et al.Formation of virus-like particles from O-type foot-and-mouth disease virus in insect cells using codon-optimized synthetic genes[J].Biotechnology Letters,2010,32(9):1223-1229.
[22] 侯立婷,陈瑾,乔绪稳,等.猪O型口蹄疫病毒细菌样颗粒疫苗的制备与免疫原性鉴定[J].生物工程学报,2017,33(2):217-227. HOU L T,CHEN J,QIAO X W,et al.Design and immunogenicity evaluation for the bacteria-like particle vaccine against swine type O foot-and-mouth disease virus[J].Chinese Journal of Biotechnology,2017,33(2):217-227.(in Chinese)
[23] MARVIN J G,MAURO P M,CHRISTOPHER S,et al.Adenovirus serotype 5 vectored foot-and-mouth disease subunit vaccines:The first decade[J].Future Virology,2010,5(1):51-64.
[24] CAROLINA S,FAYNA D S,TERESA S,et al.The pathogenesis of foot-and-mouth disease in pigs[J].Frontiers in Veterinary Science,2016,3(41):1-12.
[25] PORTA C,XU X,LOUREIRO S,et al.Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity[J].Journal of Virological Methods,2013,187(2):406-12.
[26] BHAT S A,SARAVANAN P,HOSAMANI M,et al.Novel immunogenic baculovirus expressed virus-like particles of foot-and-mouth disease (FMD) virus protect guinea pigs against challenge[J].Research in Veterinary Science,2013,95(3):1217-1223.
[27] GULLBERG M,POLACEK C,BELSHAM G J.Sequence adaptations affecting cleavage of the VP1/2A junction by the 3C protease in foot-and-mouth disease virus-infected cells[J].Journal of General Virology,2014,95(11):2402-2410.

Construction and Identification of Swine O-type Foot-and-mouth Disease Virus-like Particles

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	JIANG Haijun, WANG Dan, GUO Yu, WEI Li, HOU Lei, WANG Jing, ZHU Shanshan, LIU Jue. Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2044-2052.
[2]	LI Huawei, GUO Kewei, WANG Xuying, ZHANG Xiaoyu, LU Ziwei, LI Xinfeng, MA Hui, HOU Wenjing. Eukaryotic Expression and Subcellular Localization of Nonstructural Protein 7 of Porcine Reproductive and Respiratory Syndrome Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1160-1168.
[3]	WANG Huihui, FENG Xili, PU Feiyang, LI Yicong, WANG Mengzhu, ZHOU Xiaokai, ZHAO Zeyang, MA Zhongren, LI Zhuo, MA Xiaoxia. Prokaryotic Expression and Antigen Epitope for Non-structural Protein NS5A of BVDV Isolated from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3180-3189.
[4]	CHI Shanshan, JIA Weijuan, HE Yunjiang, MENG Qinglei, CHEN Yunjiao, WANG Xueli. Research Progress on Newcastle Disease Virus-like Particles [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1507-1515.
[5]	LI Shinian, LIU Wanning, CHEN Yaping, WANG Jintao. Design of Multi Epitope Vaccine Against Swine Acute Diarrhea Syndrome Coronavirus S, M and E Proteins Based on Immunoinformatics Methods [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1057-1066.
[6]	YANG Suzhen, SUN Yaning, LIU Yunchao, CHAI Shujun, ZHANG Gaiping. Optimization of Dual Strip for FMDV Immunized Antibody Evaluating and Discriminating Vaccinated Animals from Infected Animals [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3953-3962.
[7]	LIANG Yan, TAO Lei, SONG Xiangdong, LIU Yu, ZHANG Fengwu, LI Xuehu, XING Zhijun, LU Xihong, LIANG Jianping, FENG Jianqiang, ZHOU Xiang, WANG Zhen. Effects of ¹²C⁶⁺ Ion Beam Irradiation on the Quality of Tea Tree Essential Oil and Its Efficacy Against Foot-and-mouth Disease Virus [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 755-762.
[8]	XU Guo, LIANG Haiying, ZENG Zhiyong, WANG Bin, HUANG Tao, TANG Deyuan, DAI Zhenjiang, YE Baichuan, ZHANG Aiqiong, HE Xiaoli, XIAN Wen. Construction of Recombinant Baculovirus Expression Vector of V5-labeled Porcine Circovirus Type 2 Cap Protein [J]. , 2018, 45(7): 1775-1782.
[9]	DAI Yin, ZHANG Danjun, ZHAO Ruihong, HU Xiaomiao, SHEN Xuehuai, HOU Hongyan, PAN Xiaocheng, ZHOU Xueli. Cloning and Sequence Analysis of NS and VP Genes of Goose Parvovirus Strain in Anhui [J]. , 2018, 45(5): 1177-1183.
[10]	SUN Peng. Study on Expression and Immune Effect of F Protein of Newcastle Disease Virus Type Ⅶ in Insect Cell [J]. , 2017, 44(6): 1840-1846.
[11]	XIE San-lei, WEN Shu-xiang, LUO Lin, MA Zhi-hong, LI Gui-ping, LI Tie-liang, JIANG Na, XING Wei, SUN Hui-ling. Construction of Baculovirus Expression System and Antigenicity Analysis of Infectious Hematopoietic Necrosis Virus Glycoprotein [J]. , 2017, 44(3): 854-860.
[12]	LI Hong-jie, REN Hao-jie, LIU Yan-ke, GAO Dong-sheng, ZHAO Jun. Development and Application of a PEDV Nsp7-based Indirect ELISA for Antibody Detection [J]. , 2017, 44(11): 3306-3312.
[13]	LIU Ya-li, DING Yao-zhong, ZHANG Jie. Research Progress on the Nonstructural Proteins of Porcine Reproductive and Respiratory Syndrome Virus [J]. , 2016, 43(8): 2183-2189.
[14]	LV Shuang, YANG Tao, SUN En-cheng, XU Qing-yuan, ZHANG Ji-kai, WU Dong-lai. Subcellular Localization Analysis and Function Prediction of BTV New Type NS4 Protein [J]. , 2016, 43(1): 1-7.
[15]	GONG Zhen-li, JIANG Tao, QI Shu-yun, CHEN Guo-dong, LIU Yan-hong, LI Yong. Development of Nucleic Acid Lateral Flow Immunoassay Detection Method for Foot-and-mouth Disease Virus [J]. , 2015, 42(9): 2292-2297.