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標題: Expression and Structural Analysis of the Porcine Circovirus Type 2 Cap Protein
作者: 陳姿伃
Tzu-Yu Chen
關鍵字: 豬環狀病毒第二型
PCV2 Cap
Expression of Cap
fusion tag
引用: 張志成、黃安進、徐寶如、馬尉真、龐飛、王建雄。2011。2008至2009年台灣豬場 Torque Teno Virus (TTV) 流行概況之探討。台灣獸醫誌。37: 45-52。 Allan, G. M., Ellis, J. A., 2000. Porcine circoviruses: a review. J Vet Diagn Invest. 12, 3-14. Allan, G. M., McNeilly, F., Kennedy, S., Daft, B., Clarke, E. G., Ellis, J. A., Adair, B. M., 1998. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. J Vet Diagn Invest. 10, 3-10. Amanda L., Gavin, N. B., Hilde M. Dijstelbloem P., 1998. Identification of the mouse IgG3 receptor: implications for antibody effector function at the interface between innate and adaptive. J Immunol. 160: 20-23. Aramaki, Y., Suda, H., Tsuchiya, S., 1995. Interferon-gamma inductive effect of liposomes as an immunoadjuvant. Vaccine. 13, 1809-1814. Basta, S., Carrasco, C. P., Knoetig, S. M., Rigden, R. C., Gerber, H., Summerfield, A., McCullough, K. C., 2000. Porcine alveolar macrophages: poor accessory or effective suppressor cells for T-lymphocytes. Vet Immunol Immunopathol. 77, 177-190. Beach, N. M., Meng, X. J., 2012. Efficacy and future prospects of commercially available and experimental vaccines against porcine circovirus type 2 (PCV2). Virus Res. 164, 33-42. Beach, N. M., Ramamoorthy, S., Opriessnig, T., Wu, S. Q., Meng, X. J., 2010. Novel chimeric porcine circovirus (PCV) with the capsid gene of the emerging PCV2b subtype cloned in the genomic backbone of the non-pathogenic PCV1 is attenuated in vivo and induces protective and cross-protective immunity against PCV2b and PCV2a subtypes in pigs. Vaccine. 29, 221-232. Blanchard, P., Mahe, D., Cariolet, R., Keranflec'h, A., Baudouard, M. A., Cordioli, P., Jestin, A., 2003. Protection of swine against post-weaning multisystemic wasting syndrome (PMWS) by porcine circovirus type 2 (PCV2) proteins. Vaccine. 21, 4565-4575. Bucarey, S. A., Noriega, J., Reyes, P., Tapia, C., Saenz, L., Zuniga, A., Tobar, J. A., 2009. The optimized capsid gene of porcine circovirus type 2 expressed in yeast forms virus-like particles and elicits antibody responses in mice fed with recombinant yeast extracts. Vaccine. 27, 5781-5790. Chang, H. W., Jeng, C. R., Lin, T. L., Liu, J. J., Chiou, M. T., Tsai, Y. C., Pang, V. F., 2006. Immunopathological effects of porcine circovirus type 2 (PCV2) on swine alveolar macrophages by in vitro inoculation. Vet Immunol Immunopathol. 110, 207-219. Cheung, A. K., 2003. Transcriptional analysis of porcine circovirus type 2. Virology. 305, 168-180. Cheung, A. K., 2006. Rolling-circle replication of an animal circovirus genome in a theta-replicating bacterial plasmid in Escherichia coli. J Virol. 80, 8686-8694. Coffman, R. L., Seymour, B. W., Lebman, D. A., Hiraki, D. D., Christiansen, J. A., Shrader, B., 1988. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol Rev. 102, 5-28. Crowther, R. A., Berriman, J. A., Curran, W. L., Allan, G. M., Todd, D., 2003. Comparison of the structures of three circoviruses: chicken anemia virus, porcine circovirus type 2, and beak and feather disease virus. J Virol. 77, 13036-13041. de StGroth, S. F., & Scheidegger, D. (1980). Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 35, 1-21. Dupont, K., Nielsen, E. O., Baekbo, P., Larsen, L. E., 2008. Genomic analysis of PCV2 isolates from Danish archives and a current PMWS case-control study supports a shift in genotypes with time. Vet Microbiol. 128, 56-64. Fang, C. Y., P. Y. Lin, W. C. Ou, P. L. Chen, C. H. Shen, D. C. Chang, M. Wang., 2012. Analysis of the size of DNA packaged by the human JC virus-like particle. Journal of Virological Methods. 182, 87– 92. Fan, H., Pan, Y., Fang, L., Wang, D., Wang, S., Jiang, Y., Chen, H., Xiao, S., 2008. Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine circovirus type 2 in mice. Journal of Virological Methods. 150, 21-26. Fenaux, M., Opriessnig, T., Halbur, P. G., Elvinger, F., Meng, X. J., 2004a. A chimeric porcine circovirus (PCV) with the immunogenic capsid gene of the pathogenic PCV type 2 (PCV2) cloned into the genomic backbone of the nonpathogenic PCV1 induces protective immunity against PCV2 infection in pigs. J Virol. 78, 6297-6303. Fenaux, M., Opriessnig, T., Halbur, P. G., Elvinger, F., Meng, X. J., 2004b. Two amino acid mutations in the capsid protein of type 2 porcine circovirus (PCV2) enhanced PCV2 replication in vitro and attenuated the virus in vivo. J Virol. 78, 13440-13446. Fenaux, M., Opriessnig, T., Halbur, P. G., Meng, X. J., 2003. Immunogenicity and pathogenicity of chimeric infectious DNA clones of pathogenic porcine circovirus type 2 (PCV2) and nonpathogenic PCV1 in weanling pigs. J Virol. 77, 11232-11243. Ferrari, L., Borghetti, P., De Angelis, E., Martelli, P., 2014. Memory T cell proliferative responses and IFN-gamma productivity sustain long-lasting efficacy of a Cap-based PCV2 vaccine upon PCV2 natural infection and associated disease. Vet Res.45, 44. Finsterbusch, T., Mankertz, A., 2009. Porcine circoviruses-small but powerful. Virus Res. 143, 177-183. Finsterbusch, T., Steinfeldt, T., Doberstein, K., Rodner, C., Mankertz, A., 2009. Interaction of the replication proteins and the capsid protein of porcine circovirus type 1 and 2 with host proteins. Virology. 386, 122-131. Fort, M., Olvera, A., Sibila, M., Segales, J., Mateu, E., 2007. Detection of neutralizing antibodies in postweaning multisystemic wasting syndrome (PMWS)-affected and non-PMWS-affected pigs. Veterinary Microbiology. 125, 244–255. Fort, M., Fernandes, L. T., Nofrarias, M., Diaz, I., Sibila, M., Pujols, J., Segales, J., 2009. Development of cell-mediated immunity to porcine circovirus type 2 (PCV2) in caesarean-derived, colostrum-deprived piglets. Vet Immunol Immunopathol. 129, 101-107. Fort, M., Olvera, A., Sibila, M., Segales, J., Mateu, E., 2007. Detection of neutralizing antibodies in postweaning multisystemic wasting syndrome (PMWS)-affected and non-PMWS-affected pigs. Vet Microbiol. 125, 244-255. Fort, M., Sibila, M., Nofrarias, M., Perez-Martin, E., Olvera, A., Mateu, E., Segales, J., 2010. Porcine circovirus type 2 (PCV2) Cap and Rep proteins are involved in the development of cell-mediated immunity upon PCV2 infection. Vet Immunol Immunopathol. 137, 226-234. Fort, M., Sibila, M., Nofrarias, M., Perez-Martin, E., Olvera, A., Mateu, E., Segales, J., 2012. Evaluation of cell-mediated immune responses against porcine circovirus type 2 (PCV2) Cap and Rep proteins after vaccination with a commercial PCV2 subunit vaccine. Vet Immunol Immunopathol. 150, 128-132. Fraile, L., Sibila, M., Nofrarias, M., Lopez-Jimenez, R., Huerta, E., Llorens, A., Segales, J., 2012. Effect of sow and piglet porcine circovirus type 2 (PCV2) vaccination on piglet mortality, viraemia, antibody titre and production parameters.Vet Microbiol. 161, 229-234. Fulpius, T., Spertini, F., Reininger, L., Izui, S., 1993. Immunoglobulin heavy chain constant region determines the pathogenicity and the antigen-binding activity of rheumatoid factor. Proc Natl Acad Sci USA. 90, 2345-2349. G. M. Allan, K. V. Phenix., D. Todd, M. S. Mcnult., 1994. Some biological and physico-chemical properties of porcine circovirus. J. Vet. Med. 41, 17-26. G. W. Stevenson, M. K., S. K. Mittal, C. L. Kanitz., 1999. Ultrastructure of porcine circovirus in persistently infected PK-15 cells. Vet Pathol. 36, 368–378. Gilpin, D. F., McCullough, K., Meehan, B. M., McNeilly, F., McNair, I., Stevenson, L. S., Allan, G. M., 2003. In vitro studies on the infection and replication of porcine circovirus type 2 in cells of the porcine immune system. Vet Immunol Immunopathol. 94, 149-161. Grau-Roma, L., Fraile, L., Segales, J., 2011. Recent advances in the epidemiology, diagnosis and control of diseases caused by porcine circovirus type 2. Vet J. 187, 23-32. Greenspan, N. S., Cooper, L. J., 1992. Intermolecular cooperativity: a clue to why mice have IgG3? Immunol Today. 13, 164-168. Gu, J., Zhang, Y., Lian, X., Sun, H., Wang, J., Liu, W., Cao, R., 2012. Functional analysis of the interferon-stimulated response element of porcine circovirus type 2 and its role during viral replication in vitro and in vivo. Virol J. 9, 152. Guo, L. J., Lu, Y. H., Wei, Y. W., Huang, L. P., Liu, C. M., 2010. Porcine circovirus type 2 (PCV2): genetic variation and newly emerging genotypes in China. Virol J. 7, 273. Gurvich, O.L., Baranov, P.V., Gesteland, R.F., Atkins, J.F., 2005. Expression levels influence ribosomal frameshifting at the tandem rare arginine codons AGG_AGG and AGA_AGA in Escherichia coli. J. Bacteriol. 187, 4023-4032. Hamel, A. L., Lin, L. L., Nayar, G. P., 1998. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs. J Virol. 72, 5262-5267. Hiscox, J. A., 2002. The nucleolus-a gateway to viral infection? Arch Virol. 147, 1077-1089. Ilse Tischer, D. P., R. Rasch, S. Pociuli., 1987. Replication of porcine cireovirus induction by glueosamine and cell cycle dependence. Arch Virol. 96, 39-57. Jiang, W. and D.S. Pisetsky., 2006. The role of IFN-a and nitric oxide in the release of HMGB1 by RAW 264.7 cells stimulated with polyinosinic-polycytidylic acid or lipopolysaccharide. J Immunol. 177, 3337-3343. Kohler, G., Milstein, C., 1975. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 256, 495-497. Koonin, E. V., Ilyina, T. V., 1993. Computer-assisted dissection of rolling circle DNA replication. Biosystems. 30, 241-268. Ladekjaer-Mikkelsen, A. S., Nielsen, J., Stadejek, T., Storgaard, T., Krakowka, S., Ellis, J., Botner, A., 2002. Reproduction of postweaning multisystemic wasting syndrome (PMWS) in immunostimulated and non-immunostimulated 3-week-old piglets experimentally infected with porcine circovirus type 2 (PCV2). Vet Microbiol. 89, 97-114. Lekcharoensuk, P., Morozov, I., Paul, P. S., Thangthumniyom, N., Wajjawalku, W., Meng, X. J., 2004. Epitope mapping of the major capsid protein of type 2 porcine circovirus (PCV2) by using chimeric PCV1 and PCV2. J Virol. 78, 8135-8145. Lindblad, E. B., Elhay, M. J., Silva, R., Appelberg, R., Andersen, P., 1997. Adjuvant modulation of immune responses to tuberculosis subunit vaccines. Infect Immun. 65, 623-629. Lin, W. L., Chien, M. S., Wu, P. C., Lai, C. L., Huang, C. J., 2011. The Porcine Circovirus Type 2 Nonstructural Protein ORF3 Induces Apoptosis in Porcine Peripheral Blood Mononuclear Cells. Open Virol J. 5, 148-53. Liu, J., Bai, J., Zhang, L., Jiang, Z., Wang, X., Li, Y., Jiang, P., 2013. Hsp70 positively regulates porcine circovirus type 2 replication in vitro. Virology. 447, 52-62. Liu, J., Chen, I., Kwang, J., 2005. Characterization of a previously unidentified viral protein in porcine circovirus type 2-infected cells and its role in virus-induced apoptosis. J. Virol. 79, 8262-8274. Liu, L. J., Suzuki, T., Tsunemitsu, H., Kataoka, M., Ngata, N., Takeda, N., Li, T. C., 2008. Efficient production of type 2 porcine circovirus-like particles by a recombinant baculovirus. Arch Virol. 153, 2291-2295. Liu, Q., Tikoo, S. K., Babiuk, L. A., 2001a. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2. Virology. 285, 91-99. Liu, Q., Willson, P., Attoh-Poku, S., Babiuk, L. A., 2001b. Bacterial expression of an immunologically reactive PCV2 ORF2 fusion protein. Protein Expr Purif. 21, 115- 120. Ludwig, C., Wagner, R., 2007. Virus-like particles-universal molecular toolboxes. Curr Opin Biotechnol. 18, 537-545. Lv, Q. Z., Guo, K. K., Zhang, Y. M., 2014. Current understanding of genomic DNA of porcine circovirus type 2. Virus Genes. 49, 1-10. Eliana L. Faquim-Mauro, Robert L. Coffman, Ises A. Abrahamsohn, Mahasti S. Macedo., 1999. Mouse IgG1 antibodies comprise two functionally distinct types that are differentially regulated by IL-4 and IL-12. J Immunol. 163,3572- 3576. Eliana L. Faquim-Mauro, Mahasti S. Macedo., 2000. Induction of IL-4-dependent, anaphylactic-type and IL-4-Independent, non-anaphylactic-type IgG1 antibodies is modulated by adjuvants. International Immunology. 12, 1733–1740. Mahe, D., Blanchard, P., Truong, C., Arnauld, C., Le Cann, P., Cariolet, R., Jestin, A., 2000. Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes. J Gen Virol. 81, 1815-1824. Mankertz, A., Domingo, M., Folch, J. M., LeCann, P., Jestin, A., Segales, J., Soike, D., 2000. Characterisation of PCV-2 isolates from Spain, Germany and France. Virus Res. 66, 65-77. Mankertz, A., Hillenbrand, B., 2001. Replication of porcine circovirus type 1 requires two proteins encoded by the viral rep gene. Virology, 279, 429-438. Mankertz, A., Mueller, B., Steinfeldt, T., Schmitt, C., Finsterbusch, T., 2003. New reporter gene-based replication assay reveals exchangeability of replication factors of porcine circovirus types 1 and 2. J Virol. 77, 9885-9893. Marcekova, Z., Psikal, I., Kosinova, E., Benada, O., Sebo, P., Bumba, L., 2009. Heterologous expression of full-length capsid protein of porcine circovirus 2 in Escherichia coli and its potential use for detection of antibodies. J Virol Methods. 162, 133-141. Martelli, P., Ardigo, P., Ferrari, L., Morganti, M., De Angelis, E., Bonilauri, P., Borghetti, P., 2013. Concurrent vaccinations against PCV2 and PRRSV: study on the specific immunity and clinical protection in naturally infected pigs. Vet Microbiol. 162, 558-571. Martelli, P., Ferrari, L., Morganti, M., De Angelis, E., Bonilauri, P., Guazzetti, S., Borghetti, P., 2011. One dose of a porcine circovirus 2 subunit vaccine induces humoral and cell-mediated immunity and protects against porcine circovirus- associated disease under field conditions. Vet Microbiol. 149, 339-351. McIntosh, K. A., Harding, J. C., Ellis, J. A., Appleyard, G. D., 2006. Detection of Porcine circovirus type 2 viremia and seroconversion in naturally infected pigs in a farrow-to-finish barn. Can J Vet Res. 70, 58-61. Meehan, B. M., McNeilly, F., Todd, D., Kennedy, S., Jewhurst, V. A., Ellis, J. A., Allan, G. M., 1998. Characterization of novel circovirus DNAs associated with wasting syndromes in pigs. J Gen Virol. 79, 2171-2179. Michelle Hemann, N. M. B., Xiang-Jin Meng, Chong Wang, Patrick G. Halbur, T. O., 2014. A live-attenuated and an inactivated chimeric porcine circovirus (PCV)1-2 vaccine are both effective at inducing a humoral immune response and reducing PCV2 viremia and intrauterine infection in female swine of breeding age. The Canadian Journal of Veterinary Research. 78, 8–16. Misinzo, G., Delputte, P. L., Meerts, P., Lefebvre, D. J., Nauwynck, H. J., 2006. Porcine circovirus 2 uses heparan sulfate and chondroitin sulfate B glycosaminoglycans as receptors for its attachment to host cells. J Virol. 80, 3487-3494. N. Maclachlan, E. J. D., 2011. Fenner's Veterinary Virology. 237-242. Nawagitgul, P., Morozov, I., Bolin, S. R., Harms, P. A., Sorden, S. D., Paul, P. S., 2000. Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein. J Gen Virol. 81, 2281-2287. Nielsen, J., Vincent, I. E., Botner, A., Ladekaer-Mikkelsen, A. S., Allan, G., Summerfield, A., McCullough, K. C., 2003. Association of lymphopenia with porcine circovirus type 2 induced postweaning multisystemic wasting syndrome (PMWS). Vet Immunol Immunopathol. 92, 97-111. O'Dea, M. A., Hughes, A. P., Davies, L. J., Muhling, J., Buddle, R., Wilcox, G. E., 2008. Thermal stability of porcine circovirus type 2 in cell culture. J Virol Methods. 147, 61-66. O'Neill, K. C., Hemann, M., Gimenez-Lirola, L. G., Halbur, P. G., Opriessnig, T., 2012. Vaccination of sows reduces the prevalence of PCV-2 viraemia in their piglets under field conditions. Vet Rec. 171, 425. Opriessnig, T., O'Neill, K., Gerber, P. F., de Castro, A. M., Gimenez-Lirola, L. G., Beach, N. M., Halbur, P. G., 2013. A PCV2 vaccine based on genotype 2b is more effective than a 2a-based vaccine to protect against PCV2b or combined PCV2a/2b viremia in pigs with concurrent PCV2, PRRSV and PPV infection. Vaccine. 31, 487-494. Changsun Choi, Chanhee Chae, Edward G. Clark., 2000. Porcine postweaning multisystemic wasting syndrome in Korean pig: detection of porcine circovirus 2 infection by immunohistochemistry and polymerase chain reaction. J Vet Diagn Invest. 12, 151–153. Ramamoorthy, S., Meng, X. J., 2009. Porcine circoviruses: a minuscule yet mammoth paradox. Anim Health Res Rev. 10, 1-20. Reichert, J. M., Valge-Archer, V. E., 2007. Development trends for monoclonal antibody cancer therapeutics. Nat Rev Drug Discov. 6, 349-356. Segales, J., 2015. Best practice and future challenges for vaccination against porcine circovirus type 2. Expert Rev Vaccines. 14, 473-487. Segales, J., Allan, G. M., Domingo, M., 2005. Porcine circovirus diseases. Anim Health Res Rev. 6, 119-142. Segales, J., Kekarainen, T., Cortey, M., 2013. The natural history of porcine circovirus type 2: from an inoffensive virus to a devastating swine disease? Vet Microbiol. 165, 13-20. Segales, J., Olvera, A., Grau-Roma, L., Charreyre, C., Nauwynck, H., Larsen, L., Allan, G., 2008. PCV-2 genotype definition and nomenclature. Vet Rec. 162, 867-868. Seo, H. W., Park, C., Han, K., Chae, C., 2014. Effect of porcine circovirus type 2 (PCV2) vaccination on PCV2-viremic piglets after experimental PCV2 challenge. Vet Res. 45, 13. Shen, H. G., Zhou, J. Y., Huang, Z. Y., Guo, J. Q., Xing, G., He, J. L., Gong, L. Y., 2008. Protective immunity against porcine circovirus 2 by vaccination with ORF2- based DNA and subunit vaccines in mice. J Gen Virol. 89, 1857-1865. Sjolander, A., Lovgren Bengtsson, K., Johansson, M., Morein, B., 1996. Kinetics, localization and isotype profile of antibody responses to immune stimulating complexes (iscoms) containing human influenza virus envelope glycoproteins. Scand J Immunol. 43, 164-172. Stevens, T. L., Bossie, A., Sanders, V. M., Fernandez-Botran, R., Coffman, R. L., Mosmann, T. R., Vitetta, E. S., 1988. Regulation of antibody isotype secretion by subsets of antigen-specific helper T cells. Nature. 334, 255-258. Streeck, H., Frahm, N., Walker, B. D., 2009. The role of IFN-g Elispot assay in HIV vaccine research. Nature Protocols. 4, 461-469. Timmusk, S., Fossum, C., Berg, M., 2006. Porcine circovirus type 2 replicase binds the capsid protein and an intermediate filament-like protein. J Gen Virol. 87, 3215- 3223. Trundova, M., Celer, V., 2007. Expression of porcine circovirus 2 ORF2 gene requires codon optimized E. coli cells. Virus Genes. 34, 199-204. Truong, C., Mahé, D., Blanchard, P., Le Dimna, M., Madec, F., Jestin, A., Albina, E., 2001. Identification of an immunorelevant ORF2 epitope from porcine circovirus type 2 as a serological marker for experimental and natural infection. Arch. Virol. 146, 1197-1211. Washington, D., 1999. Committee on methods of producing monoclonal antibodies, institute for laboratory Animal Research, National Research Council. ISBN: 0-309-51904-7 Walker, I.W., Konoby, C.A., Jewhurst, V.A., McNair, I., McNeilly, F., Meehan, B.M., Cottrell, T.S., Ellis, J.A., Allan, G.M., 2000. Development and application of a competitive enzyme-linked immunosorbent assay for the detection of serum antibodies to porcine circovirus type 2. J. Vet. Diagn. Invest. 12, 400-405. Yin, S., Sun, S., Yang, S., Shang, Y., Cai, X., Liu, X., 2010. Self-assembly of virus-like particles of porcine circovirus type 2 capsid protein expressed from Escherichia coli. Virol J. 7, 166. Yoichi Takahagi, S. T., Yasutaka nishiyama, Fumiki morimatsu, Hiroshi murakami., 2010. Differential effects of porcine circovirus type 2 (PCV2) vaccination on PCV2 genotypes at Japanese pig farms. J. Vet. Med. Sci. 72, 35–41. Zhang S.P., Zubay G., Goldman E., 1991. Low-usage codons in Escherichia coli, yeast, fruit fly and primates. Gene 105, 61-72. Zhai, S. L., Chen, S. N., Xu, Z. H., Tang, M. H., Wang, F. G., Li, X. J., Wei, W. K., 2014. Porcine circovirus type 2 in China: an update on and insights to its prevalence and control. Virol J. 11, 88. Zhou, J. Y., Shang, S. B., Gong, H., Chen, Q. X., Wu, J. X., Shen, H. G., Guo, J. Q., 2005. In vitro expression, monoclonal antibody and bioactivity for capsid protein of porcine circovirus type II without nuclear localization signal. J Biotechnol. 118, 201-211.
摘要: Porcine circovirus type 2 (PCV2) is the primary causative agent of the economically important global disease, porcine circovirus associated diseases (PCVAD). The viral capsid protein (Cap) containing the major antigenic sites has become the target protein for developing of diagnostic reagents and PCV2 subunit vaccine. The purpose of this study is to express a variety of recombinant Cap proteins using an E. coli expression system for further analysis of the structure and immunogenicity of viral capsid. Four recombinant Cap proteins including the rCapT7+ containing a N-terminal T7 tag, the rCapHis+ containing a C-terminal His tag, the rCapT7+His+ containing both tags at each end, and the rCap containing no tag were constructed. The recombinant Cap proteins were purified by the continuous sucrose gradient ultracentrifugation and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All the expressed Cap proteins no matter containing a fusion tag could self-assemble into virus-like particles (VLPs) by the transmission electronmicroscope analysis. The immunogenicity of PCV2 VLPs were further evaluated in mice. Both the rCap and fusion tags containing rCapT7+His+ VLPs immunized groups were able to elicit specific antibody responses to PCV2 with neutralizing antibody titers of 1:8 after twice booster immunization. In addition, both immunized groups showed detectable of IFN- secreting cells response in the ELISpot analysis. All the results demonstrated that the PCV2 VLPs based on the E. coli-expressed Cap proteins could mount both humoral and cellular immune response against PCV2 Cap in mice, and the terminal fusion tags had no significant effect on structural assembly and immunogenicity of Cap protein.
文章公開時間: 2018-06-22
Appears in Collections:微生物暨公共衛生學研究所



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