犬瘟热病毒囊膜糖蛋白在杆状病毒-昆虫细胞系统中的表达及鉴定

doi:10.11843/j.issn.0366-6964.2015.06.014

Abstract

Abstract:

The objectives of this study were to express the native structure of fusion protein (F) and hemagglutinin glycoprotein (H) of canine distemper virus (CDV).The F and H gene of lesser panda attenuated strain were amplified by PCR and cloned into pFastBac^TM1 vector.The recombinant plasmids were sequenced，and then were transformed into competent DH10Bac^TM E.coli cells to construct shuttle plasmids，rBacmid F and rBacmid H，by homologous recombination.The recombinant plasmids were then transfected into Sf9 cells to construct recombinant baculoviruses and the expression products of rF and rH were identified with IFA and Western blot.The expression of rF and rH in insect cells infected with recombinant baculoviruses were identified by IFA with dog anti-CDV hyperimmune serum and were detected by Western blot with mouse polyclonal antibody against F and H major epitopes of CDV.The molecular weight of expressed F and H protein were identified as 63 and 68 kD，which were consistent with the expected value.The results show that both envelope glycoproteins are successfully expressed in baculovirus-insect cell expression system and have a good immunoreactivity.Our study laid the foundation for the development of the CDV virus-like particle vaccine.

CLC Number:

S852.659.5

YU Yi-cong，FENG Na，YAN Fei-hu，GAI Wei-wei，WANG Tie-cheng，WANG Hua-lei，ZHENG Xue-xing，ZHAO Yong-kun，HUANG Geng，YANG Song-tao，GAO Yu-wei，XIA Xian-zhu. Expression and Identification of Glycoprotein of Canine Distemper Virus in the Baculovirus-Insect Cell System[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(6): 981-988.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2015.06.014

https://www.xmsyxb.com/EN/Y2015/V46/I6/981

References

［1］WANG F，YAN X，CHAI X，et al.Differentiation of canine distemper virus isolates in fur animals from various vaccine strains by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism according to phylogenetic relations in China［J］.Virol J，2011，8：85.
［2］蔡宝祥.家畜传染病学［M］.2版.北京：中国农业出版社，1999：301-304.
CAI B X.Livestock infectious disease ［M］.2ed edition.Beijing：China Agriculture Press，1999：301-304.(in Chinese)
［3］APPEL M J G，MONTALI R J.Canine distemper and emerging morbillivirus diseases in exotic species［C］.Proc Am Assoc Zoo Vet，1994：336-339.
［4］李金中，夏咸柱，邱薇，等.小熊猫等四种动物犬瘟热病毒的分离与鉴定［J］.中国兽医科技，1998，28(8)：8-10.
LI J Z，XIA X Z,QIU W,et al.Isolation and identification of canine distemper virus from lesser panda，dog，raccoon dog and fox［J］.Chinese Journal of Veterinary Science and Technology，1998，28(8)：8-10.(in Chinese)
［5］胡桂学，夏咸柱，李金中.大熊猫犬瘟热致死性感染的首次报告［C］.中国微生物学会兽医微生物学学术年会论文集.北京，1997，1(997)：1.
HU G X，XIA X Z，LI J Z.The first report of giant panda infected with lethal canine distemper［C］.Chinese Society for Microbiology Veterinary Microbiology Academic Conference.Beijing，1997，1(997)：1.(in Chinese)
［6］QIU W，ZHENG Y，ZHANG S，et al.Canine distemper outbreak in rhesus monkeys，China［J］.Emerg Infect Dis，2011，17(8)：1541-1543.
［7］NORRIS J M，KROCKENBERGER M B，BAIRD A A，et al.Canine distemper：re-emergence of an old enemy［J］.Aust Vet J，2006，84(10)：362-363.
［8］殷震，刘景华.动物病毒学［M］.2版.北京：科学出版社，1997：756-762.
YIN Z，LIU J H.Animal virology［M］.2ed edition.Beijing：Science Press，1997：756-762.(in Chinese)
［9］VON MESSLING V，ZIMMER G，HERRLER G，et al.The hemagglutinin of canine distemper virus determines tropism and cytopathogenicity［J］.J Virol，2001，75(14)：6418-6427.
［10］潘耀谦.犬瘟热发病机理的免疫及分子病理学研究——脑组织及淋巴器官病变观察［D］.北京：中国农业大学，2005.
PAN Y Q.Studies of pathogenesis of immune and molecular pathology on dogs with canine distemper—pathologic observation in brain tissues and lymphatic tissues［D］.Beijing：China Agricultural University，2005.(in Chinese)
［11］萨姆布鲁克 J，弗里奇 E F，曼尼阿蒂斯 T.分子克隆实验指南［M］.2版.北京：科学出版社，1993：674-683.
SAMBMOK J，FRITSEH E F，MANIATIS T.Molecular cloning a laboratory manual［M］.2ed edition.Beijing：Science Press，1993：674-683.(in Chinese)
［12］DEEM S L，SPELMAN L H，YATES R A，et al.Canine distemper in terrestrial carnivores：a review［J］.J Zoo Wildl Med，2000，31(4)：441-451.
［13］MARTELLA V，ELIA G，BUONAVOGLIA C.Canine distemper virus［J］.Vet Clin North Am Small Anim Pract，2008，38(4)：787-797.
［14］孔庆波.犬瘟热免疫预防研究进展［J］.动物医学进展，2007，28(6)：69-75.
KONG Q B.Progress on immune prevention of canine distemper［J］.Progress in Veterinary Medicine，2007，28(6)：69-75.(in Chinese)
［15］BARRETT T.Morbillivirus infections，with special emphasis on morbilliviruses of carnivores［J］.Vet Microbiol，1999，69(1-2)：3-13.
［16］王磊，李天松，刘玉秀，等.犬瘟热新型疫苗研究进展［J］.动物医学进展，2011，32(3)：99-103.
WANG L，LI T S，LIU Y X，et al.Progress on new type vaccines of canine distemper［J］.Progress in Veterinary Medicine，2011，32(3)：99-103.(in Chinese)
［17］WELTER J，TAYLOR J，TARTAGLIA J，et al.Vaccination against canine distemper virus infection in infant ferrets with and without maternal antibody protection，using recombinant attenuated poxvirus vaccines［J］.J Virol，2000，74(14)：6358-6367.
［18］高炳淼，李宝珠，于津鹏，等.外源基因在昆虫杆状病毒表达系统中的表达［J］.中国生物工程杂志，2011，31(11)：123-129.
GAO B M，LI B Z，YU J P，et al.Expression of foreign genes in insect baculovirus expression systems［J］. China Biotechnology，2011，31(11)：123-129.(in Chinese)
［19］AILOR E，BETENBAUGH M J.Modifying secretion and post-translational processing in insect cells［J］.Curr Opin Biotechnol，1999，10(2)：142-145.
［20］DAI X，HAJÓS J P，JOOSTEN N N，et al.Isolation of a Spodoptera exigua baculovirus recombinant with a 10.6 kbp genome deletion that retains biological activity［J］.J Gen Virol，2000，81(10)：2545-2554.
［21］SANDOVAL C，CURTIS H，CONGOTE L F.Enhanced proliferative effects of a baculovirus-produced fusion protein of insulin-like growth factor and α1-proteinase inhibitor and improved anti-elastase activity of the inhibitor with glutamate at position 351［J］.Protein Eng，2002，15(5)：413-418.
［22］FRAZER I H.Cervical cancer vaccine development［J］.Sex Health，2010，7(3)：230-234.
［23］MENA J A，KAMEN A A.Insect cell technology is a versatile and robust vaccine manufacturing platform［J］.Expert Rev Vaccines，2011，10(7)：1063-1081.
［24］全传松，姜骞，于作，等.犬瘟热病毒血凝蛋白在昆虫细胞中的表达与抗原性分析［J］.中国畜牧兽医，2014，41(8)：25-29.
QUAN C S，JIANG Q，YU Z，et al.Expression of hemagglutinin of canine distemper virus in the insect cell and its antigenic analysis［J］.China Animal Husbandry & Veterinary Medicine，2014，41(8)：25-29.(in Chinese)
［25］隋修锟，金红岩，李文超，等.小反刍兽疫病毒H基因在杆状病毒中的表达及其免疫原性研究［J］.畜牧兽医学报，2014，45(6)：974-980.
SUI X K，JIN H Y，LI W C，et al.Expression of H gene of Peste des Petits Ruminants virus in baculovirus and its immunogenicity study［J］.Acta Veterinaria et Zootechnica Sinica，2014，45(6)：974-980.(in Chinese)
［26］冯佩平.犬瘟热病毒主要结构蛋白基因在昆虫细胞中的表达及间接 ELISA 检测方法的建立［D］.长春：吉林农业大学，2012.
FENG P P.Expression of CDV major structural protein in Sf9 cells and establishment of indirect ELISA assay［D］.Changchun：Jilin Agricultural University，2012.(in Chinese)
［27］刘拂晓.小反刍兽疫病毒样颗粒的构建及对小鼠免疫效力的评价［D］.长春：吉林大学，2013.
LIU F X.Construction of Peste des Petits Ruminants virus-like particles and evaluation of their immune effects in mice［D］.Changchun：Jilin University，2013.(in Chinese)
［28］LI W，JIN H，SUI X，et al.Self-assembly and release of peste des petits ruminants virus-like particles in an insect cell-baculovirus system and their immunogenicity in mice and goats［J］.PLoS One，2014，9(8)：e104791.

[1]	MENG Lingzhai, CHEN Chunli, YU Mengmeng, WANG Zhanxin, WANG Suyan, LIU Peng, HE Tana, GUO Ru, CHEN Yuntong, LIU Changjun, QI Xiaole, WU Zhiqiang, GAO Yulong. Pathogenicity of Avian Metapneumovirus Subtype B on Yellow Feather Broilers and Evaluation on Immune Effect of Inactivated Vaccine [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5154-5161.
[2]	JIANG Shengqiang, HU Jing, CHEN Hongying. Expression Analysis of CircRNAs in A549 Cells Infected with H1N1 Influenza A Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4724-4734.
[3]	ZHANG Ao, TAN Bin, LIU Kexin, LIU Jiali, ZHANG Shuqin. Genomic Characterization Analysis of a H1N1 Subtype Swine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4866-4871.
[4]	HE Chenxiang, GAO Shandian, TIAN Zhancheng, DU Junzheng, WANG Jinming, GUAN Guiquan, YIN Hong. Development of a Differential Diagnostic ELISA Based on G_NS Protein to Distinguish BEFV Infected and Vaccinated Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4320-4326.
[5]	CHANG Yiming, TANG Cheng, YUE Hua. Molecular Epidemiological Investigation of Bovine Respiratory Syncytial Virus in Yaks [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2030-2041.
[6]	FANG Yuan, HOU Qiaodi, XIANG Chaohui, ZHAO Hongyi, QI Xuefeng. Regulatory Effects of IFITM3 on Proliferation of Peste des Petits Ruminants Virus (PPRV) in Goat Endometrial Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2200-2207.
[7]	LI Shun, LI Guizhen, YUAN Yaoxian, LI Kangjian, MA Chunquan, LI Shoujun, HUANG Liangzong, MA Jun. Effect of the Length of PA-X Gene on Replication and Pathogenicity of H3N2 Canine Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 272-280.
[8]	CUI Mingxian, WANG Xingbo, HUANG Yanming, BIAN Xiyi, FENG Mengke, YAN Yan, DONG Weiren, ZHOU Jiyong. Genetic Characterization and Evolution of Three Strains of H3N2 Avian Influenza Viruses [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4116-4122.
[9]	YU Mengmeng, BAO Yuanling, WANG Suyan, XIN Ziqi, LIU Peng, FENG Xiaoyan, MENG Lingzhai, GUO Ru, ZHANG Yanping, LIU Changjun, QI Xiaole, LI Junping, WANG Xiaomei, GAO Yulong. Isolation, Identification and Pathogenicity of Subtype B Avian Metapneumovirus [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3540-3549.
[10]	ZHOU Yong, LI Zhixin, LU Hongwei, SUN Yan, LI Tian, DU Fanshu, PU Juan. Surveillance and Outbreak Analysis of H5 and H7N9 Subtypes of Highly Pathogenic Avian Influenza in China [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3093-3106.
[11]	PAN Chunrong, DENG Ruixue, HU Linjie, ZHU Xueliang, SUN Yuefeng, WANG Guirong, MENG Xuelian. Effect of Histone Deacetylase Inhibitors on Peste Des Petits Ruminants Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2307-2316.
[12]	CHEN Zixuan, ZHANG Nan, HU Qun, QUAN Keji, QIN Tao, CHEN Sujuan, PENG Daxin, LIU Xiufan. Mutation Analysis of Antigen Sites 145 and 153 on Hemagglutinin of H9N2 Avian Influenza Virus in China [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1165-1172.
[13]	DUAN Zhiqiang, XIE Lingling, CHEN Jiaqi, TANG Hong, WANG Yanbi, ZHAO Caiqin, ZHAO Jiafu. Research Progress on the Interactions of Newcastle Disease Virus M Protein with Host Proteins [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 32-42.
[14]	YAN Yayao, GU Min, LIU Xiufan. Advance in the Influence of Amino Acid Variation in HA Protein on the Biological Properties of H7N9 Subtype Influenza Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2093-2106.
[15]	JIANG Mei, CHEN Wu, ZHAI Junqiong, BU Wandi, XIE Yilun, LIU Canbin, SHAN Fen, LUO Manlin. Cloning and Sequence Analysis of H and F Genes of Canine Distemper Virus Strain from Red Panda [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1670-1676.

Expression and Identification of Glycoprotein of Canine Distemper Virus in the Baculovirus-Insect Cell System

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments