AMB  >> Vol. 6 No. 3 (September 2017)

    Display of Classical Swine Fever Virus E2 Protein BC Antigenic Domain on the Cell Surface of Saccharomyces cerevisiae

  • 全文下载: PDF(1254KB) HTML   XML   PP.72-78   DOI: 10.12677/AMB.2017.63010  
  • 下载量: 215  浏览量: 787   国家自然科学基金支持


刘小凤,汪 倩,罗明阳,孙金福:东北大学生命科学与健康学院生物技术研究所,辽宁 沈阳

酵母表面展示猪瘟病毒E2BC抗原域口服疫苗Yeast Surface Display Classical Swine Fever Virus E2BC Antigenic Domain Oral Vaccine


为构建猪瘟病毒(CSFV)E2蛋白的B、C抗原域(E2BC)酿酒酵母表面展示体系,以pVEXE2为模板,用PCR克隆了E2BC基因,并插入酵母展示表达载体p1v5AG的BamH I和EcoRI位点,构建重组酵母展示表达载体,用LiAC方法转化酿酒酵母细胞W303,构建重组酵母菌。重组菌经培养后,对酵母菌细胞进行针对His标签的间接免疫荧光染色,检测His-E2BC融合表达蛋白。结果显示,重组酵母菌表面有绿色荧光,表明E2BC蛋白成功表达于酵母表面,为开发酵母载体口服疫苗奠定了基础。

To construct the yeast display system of classical swine fever virus E2 glycoprotein BC antigenic domain, gene fragment was cloned by PCR using pVEXE2 as a template. E2BC was inserted into the site of BamH I and EcoR I of p1v5AG to produce recombinant surface displaying vector of p1E2BCAG. p1E2BCAG was transformed into Saccharomyces cerevisiae by LiAC method. After recombinant cells were cultivated, indirect immunofluorescence was used to detect E2BC expression. The results indicate that E2BC protein was displayed on the surface of Saccharomyces cerevisiae cell successfully. This work provides a foundation for the development of Saccharomyces cerevisiae vector oral vaccine of CSFV E2BC protein.

刘小凤, 汪倩, 罗明阳, 孙金福. 猪瘟病毒E2蛋白BC抗原域的酿酒酵母表面展示[J]. 微生物前沿, 2017, 6(3): 72-78.


[1] Simmonds, P., Becher, P., Collett, M.S., et al. (2011) Family Flaviridae. In: King, A.M.Q., Lefkowitz, E. and Adams, M.J., Eds., Virus Taxonomy. Eighth Report of the International Committee on Taxonomy of Virus, Academic Press, San Diego, 1002-1020.
[2] Lindenbach, B.D., Thiel, H.J. and Rice, C.M. (2007) Flaviviridae: The Viruses and Their Replication. 5th Edition, Lippincott Williams & Wilkins, Philadelphia, PA.
[3] Van Rijn, P.A., Van Gennip, H.G., de Meijer, E.J. and Moormann, R.J.M. (1993) Epitope Mapping of Envelope Glycoprotein E1 of Hog Cholera Virus Strain Brescia. Journal of General Virology, 74, 2053-2060.
[4] Van Rijn, P.A., Miedema, G.K., Wensvoort, G. and Moormann, R.J.M. (1994) Antigenic Structure of Envelope Glycoprotien E1 of Hog Cholera Virus. Journal of Virology, 68, 3934-3942.
[5] 郭钦, 张伟, 阮晖, 等. 酿酒酵母表面展示表达系统及应用[J]. 中国生物工程杂志, 2008, 28(12): 116-122.
[6] Vinopal, S., Ruml, T. and Kotrba, P. (2007) Biosorption of Cd2+ and Zn2+ by Cell Surface-Engineered Saccharomyces cerevisiae. International Biodeterioration & Biodegradation, 60, 96-102.
[7] Gietz, R.D., Schiestl, R.H., Willems, A.R. and Woods, R.A. (1995) Studies on the Transformation of Intact Yeast Cells by the LiAc/SS-DNA/PEG Procedure. Yeast, 11, 355-360.
[8] 向柱方, 林影, 叶波, 等. HIV-1 gp41的酵母表面展示及表达优化[J]. 生物工程学报, 2008, 24(4): 684-689.
[9] Stubbs, A.C., Martin, K.S., Coeshott, C., et al. (2001) Whole Recombinant Yeast Vaccine Activates Dendritic Cells and Elicits Protective Cell-Mediated Immunity. Nature Medicine, 7, 625-629.
[10] Wasilenko, J.L., Sarmento, L., Spatz, S. and Pantin-Jackwood, M. (2010) Cell Surface Display of Highly Pathogenic Avian Influenza Virus Hemagglutinin on the Surface of Pichia pastoris Cells Using Alpha-Agglutinin for Production of Oral Vaccines. Biotechnology Progress, 26, 542-547.
[11] Lei, H., Jin, S., Karlsson, E., et al. (2016) Yeast Surface-Displayed H5N1 Avian Influenza Vaccines. Journal of Immunology Research, 2016, 4131324.
[12] Kim, J.M., Jung, D.I., Eom, Y.J., Park, S.M., Yoo, H.S., Jang, Y.S., Yang, M.S. and Kim, D.H. (2010) Surface-Displayed Expression of a Neutralizing Epitope of ApxIIA Exotoxin in Saccharomyces cerevisiae and Oral Administration of It for Protective Immune Responses against Challenge by Actinobacillus pleuropneumoniae. Bioscience, Biotechnology, and Biochemistry, 74, 1362-1367.
[13] Zhao, J.Z., Xu, L.M., Liu, M., et al. (2017) Preliminary Study of an Oral Vaccine against Infectious Hematopoietic Necrosis Virus Using Improved Yeast Surface Display Technology. Molecular Immunology, 85, 196-204.
[14] Dong, X.N., Chen, Y., Wu, Y. and Chen, Y.H. (2005) Candidate Mul-ti-Peptide-Vaccine against Classical Swime Fever Virus Induced Potent Immunity with Serological Marker. Vaccine, 23, 3630-3633.