Expression and functional analysis of Brucella outer membrane protein 25 in recombinant goat pox virus

Sun,Zhihua; Liu,Laizhen; Zhang,Hui; Li,Youwen; Wei,Feng; Li,Zhiqiang; Wang,Pengyan; Fu,Qiang; Ren,Yan; Zhang,Yu; Guo,Zhiru; Chen,Chuangfu

doi:10.3892/mmr.2019.9868

Expression and functional analysis of Brucella outer membrane protein 25 in recombinant goat pox virus

Authors:
- Zhihua Sun
- Laizhen Liu
- Hui Zhang
- Youwen Li
- Feng Wei
- Zhiqiang Li
- Pengyan Wang
- Qiang Fu
- Yan Ren
- Yu Zhang
- Zhiru Guo
- Chuangfu Chen
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Affiliations: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, P.R. China, College of Biology and Food, Shangqiu Normal University, Shangqiu, Henan 476000, P.R. China, College of Medicine, Shihezi University, Shihezi, Xinjiang 832000, P.R. China
Published online on: January 16, 2019 https://doi.org/10.3892/mmr.2019.9868
Pages: 2323-2329

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Abstract

The Capripoxvirus (CaPV) has a large double‑stranded DNA genome and a restricted host‑range. At present, it is being investigated as an ideal vaccine vector. In the present study, a novel recombinant goat pox virus (rGTPV) was constructed to express Brucella outer membrane protein (OMP)25, and was validated by in vitro and in vivo immunization assays. A novel rGTPV vector was created, in which the thymidine kinase gene was used as a flanking sequence, I1L was inserted as a promoter element to enhance Brucella OMP25 expression, and p7.5 as another promoter element was used to regulate guanine phosphoribosyl‑transferase as a selection maker. The rGTPV vector was transfected into sheep fetal fibroblast/lamb testis cells pre‑infected with GTPV G14‑STV44‑55 to recombine. Brucella OMP25 protein was expressed in cells by rGTPV, and activated immune reactivity to Brucella OMP25 protein, as detected by western blotting. Furthermore, rGTPV elicited, anti‑Brucella‑specific immunoglobulin G responses, as determined by ELISA. Mice vaccinated with rGTPV did not exhibit pathology alterations in the kidney and liver. These results suggested that the novel rGTPV was able to efﬁciently drive Brucella OMP25 protein expression and activate immune reactivity, and may have applications in CaPV live vector vaccines and associated research.

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1	Bercovich Z: The use of skin delayed-type hypersensitivity as an adjunct test to diagnose brucellosis in cattle: A review. Vet Q. 22:123–130. 2000. View Article : Google Scholar : PubMed/NCBI
2	Wang Z and Wu Q: Research progress in live attenuated Brucella vaccine development. Curr Pharm Biotechnol. 14:887–896. 2013. View Article : Google Scholar : PubMed/NCBI
3	Cloeckaert A, de Wergifosse P, Dubray G and Limet J: Identification of seven surface-exposed Brucella outer membrane proteins by use of monoclonal antibodies: Immunogold labeling for electron microscopy and enzyme-linked immunosorbent assay. Infect Immun. 58:3980–3987. 1990.PubMed/NCBI
4	Shevtsov A, Tarlykov P, Zholdybayeva E, Shevtsova E, Momynkulov D, Sytnik I, Karibaev T, Chsherbakov A and Momynaliev K: Draft genome sequence of the live vaccine strain Brucella abortus 82. Genome Announc. 1:e01101–01113. 2013. View Article : Google Scholar : PubMed/NCBI
5	Schurig GG, Sriranganathan N and Corbel MJ: Brucellosis vaccines: Past, present and future. Vet Microbiol. 90:479–496. 2002. View Article : Google Scholar : PubMed/NCBI
6	Diallo A, Minet C, Berhe G, Le Goff C, Black DN, Fleming M, Barrett T, Grillet C and Libeau G: Goat immune response to capripox vaccine expressing the hemagglutinin protein of peste des petits ruminants. Ann N Y Acad Sci. 969:88–91. 2002. View Article : Google Scholar : PubMed/NCBI
7	Venkatesan G, Balamurugan V, Yogisharadhya R, Kumar A and Bhanuprakash V: Differentiation of sheeppox and goatpox viruses by polymerase Chain reaction-restriction fragment length polymorphism. Virol Sin. 27:353–359. 2012. View Article : Google Scholar : PubMed/NCBI
8	Zhao Z, Wu G, Zhu X, Yan X, Dou Y, Li J, Zhu H, Zhang Q and Cai X: RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells. Virol J. 9:482012. View Article : Google Scholar : PubMed/NCBI
9	Mackett M, Smith GL and Moss B: Vaccinia virus: A selectable eukaryotic cloning and expression vector. Proc Natl Acad Sci USA. 79:7415–7419. 1982. View Article : Google Scholar : PubMed/NCBI
10	Liu X, Kremer M and Broyles SS: A natural vaccinia virus promoter with exceptional capacity to direct protein synthesis. J Virol Methods. 122:141–145. 2004. View Article : Google Scholar : PubMed/NCBI
11	Falkner FG and Moss B: Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors. J Virol. 62:1849–1854. 1988.PubMed/NCBI
12	Fick WC and Viljoen GJ: Identification and characterisation of an early/late bi-directional promoter of the capripoxvirus, lumpy skin disease virus. Arch Virol. 144:1229–1239. 1999. View Article : Google Scholar : PubMed/NCBI
13	Chen H, Li Y, Li Z, Shi J, Shinya K, Deng G, Qi Q, Tian G, Fan S, Zhao H, et al: Properties and dissemination of H5N1 viruses isolated during an influenza outbreak in migratory waterfowl in western China. J Virol. 80:5976–5983. 2006. View Article : Google Scholar : PubMed/NCBI
14	Feng W, Zhiru G, Chuangfu C and Jun Q: Construction of the recombinant goat pox virus expressing OMP 25 gene of B. melitensis. Prog Veterinary Med. 29:6–9. 2008.
15	Goel D, Rajendran V, Ghosh PC and Bhatnagar R: Cell mediated immune response after challenge in Omp25 liposome immunized mice contributes to protection against virulent Brucella abortus 544. Vaccine. 31:1231–1237. 2013. View Article : Google Scholar : PubMed/NCBI
16	Gershon PD and Black DN: A comparison of the genomes of capripoxvirus isolates of sheep, goats, and cattle. Virology. 164:341–349. 1988. View Article : Google Scholar : PubMed/NCBI
17	Tulman ER, Afonso CL, Lu Z, Zsak L, Sur JH, Sandybaev NT, Kerembekova UZ, Zaitsev VL, Kutish GF and Rock DL: The genomes of sheeppox and goatpox viruses. J Virol. 76:6054–6061. 2002. View Article : Google Scholar : PubMed/NCBI
18	Berhe G, Minet C, Le Goff C, Barrett T, Ngangnou A, Grillet C, Libeau G, Fleming M, Black DN and Diallo A: Development of a dual recombinant vaccine to protect small ruminants against peste-des-petits-ruminants virus and capripoxvirus infections. J Virol. 77:1571–1577. 2003. View Article : Google Scholar : PubMed/NCBI