• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

Message Board

Respected readers, authors and reviewers, you can add comments to this page on any questions about the contribution, review,        editing and publication of this journal. We will give you an answer as soon as possible. Thank you for your support!

Name
E-mail
Phone
Title
Content
Verification Code
Volume 33 Issue 9
Mar.  2019
Turn off MathJax
Article Contents
PAN Hui, ZHAO Zhong-hao, ZHANG Song-bai, ZHANG De-yong, ZHANG Jie, LI Jun-kai, ZHENG Li-min. Preparation and Application of Polyclonal Antibodies Against Nucleocapsid Protein N and Envelope Membrane Glycoprotein Gn of Tomato Spotted Wilt Orthotospovirus[J]. Fujian Journal of Agricultural Sciences, 2018, 33(9): 963-968. doi: 10.19303/j.issn.1008-0384.2018.09.013
Citation: PAN Hui, ZHAO Zhong-hao, ZHANG Song-bai, ZHANG De-yong, ZHANG Jie, LI Jun-kai, ZHENG Li-min. Preparation and Application of Polyclonal Antibodies Against Nucleocapsid Protein N and Envelope Membrane Glycoprotein Gn of Tomato Spotted Wilt Orthotospovirus[J]. Fujian Journal of Agricultural Sciences, 2018, 33(9): 963-968. doi: 10.19303/j.issn.1008-0384.2018.09.013

Preparation and Application of Polyclonal Antibodies Against Nucleocapsid Protein N and Envelope Membrane Glycoprotein Gn of Tomato Spotted Wilt Orthotospovirus

doi: 10.19303/j.issn.1008-0384.2018.09.013
  • Received Date: 2018-05-18
  • Rev Recd Date: 2018-08-20
  • Publish Date: 2018-09-01
  • Tomato spotted wilt disease caused by tomato spotted wilt orthotospovirus (TSWV) has resulted seriously yield and quality losses in vegetables and flowers in China. In this study, nucleocapsid protein N gene and membrane polysaccharide protein Gn gene of TSWV were cloned into prokaryotic expression vectors, respectively. The plasmids were transformed into Rosetta(DE3) cells for expression of N or Gn proteins, which were used as immunogens to immunize New Zealand white rabbits, then the polyclonal antibodies against N and Gn proteins were successfully prepared. Western blot analysis showed that the polyclonal antibody against N protein could bind to the N protein specifically. Indirect ELISA showed that the titer of antiserum against N protein was about 1:12 800. However, the specificity of antibody against Gn protein was poor, and the titer was not detected. The polyclonal antibody against N protein has been prepared successfully, and it can be a useful reagent for disease diagnosis in field, the research of virus localization in insect body and the interaction of virus and insect vector.
  • loading
  • [1]
    PARRELLA G, GOGNALONS P, GEBRE-SELASSIè K, et al. An update of the host range of tomato spotted wilt virus[J]. Journal of Plant Pathology, 2003, 85(4):227-264. http://www.jstor.org/stable/41998156
    [2]
    CHO J J, RFL M, GERMAN T L, et al. A multidisciplinary approach to management of tomato spotted wilt virus in Hawaii[J]. Plant Disease, 1989, 73(5):375-383. doi: 10.1094-PD-73-0375/
    [3]
    SCHOLTHOF K B, ADKINS S, CZOSNEK H, et al. Top 10 plant viruses in molecular plant pathology[J]. Molecular Plant Pathology, 2011, 12(9):938-954. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=6fcee62f0c982d24b26117a9fd857d06
    [4]
    李洁, 迟胜起, 杨勤民, 等.山东烟台地区发生番茄斑萎病毒病危害[J].植物保护, 2017, 43(1):228-232. doi: 10.3969/j.issn.0529-1542.2017.01.043
    [5]
    张仲凯.番茄斑萎病毒属病毒粒体在寄主植物中的装配与运动机制研究[D].北京: 中国农业科学院, 2015.
    [6]
    KING A M Q, CARSTENS E B, LEFKOWITZ E J. Ninth report of the international Committee on taxonomy of viruses[J]. Elsevier, 2011:774-796.
    [7]
    DE H P, KORMELINK R, DE O R R, et al. Tomato spotted wilt virus L RNA eNCodes a putative RNA polymerase[J]. Journal of General Virology, 1991, 72(9):2207-2216. doi: 10.1099/0022-1317-72-9-2207
    [8]
    VAN K I, GOLDBACH R, KORMELINK R. Purified tomato spotted wilt virus particles support both genome replication and transcription in vitro[J]. Virology, 2002, 303(2):278. http://europepmc.org/abstract/MED/12490389
    [9]
    KORMELINK R, STORMS M, VAN L J, et al. Expression and subcellular location of the NSM protein of tomato spotted wilt virus (TSWV), a putative viral movement protein[J]. Virology, 1994, 200(1):56-65. doi: 10.1006/viro.1994.1162
    [10]
    STORMS M M, KORMELINK R, PETERS D, et al. The nonstructural NSm protein of tomato spotted wilt virus induces tubular structures in plant and insect cells[J]. Virology, 1995, 214(2):485-493. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=4a7fe9353a0ae3c440853c1b95c1fccb
    [11]
    LEWANDOWSKI D J, ADKINS S. The tubule-forming NSm protein from Tomato spotted wilt virus complements cell-to-cell and long-distaNCe movement of Tobacco mosaic virus hybrids[J]. Virology, 2005, 342(1):26. doi: 10.1016/j.virol.2005.06.050
    [12]
    LI W, LEWANDOWSKI D J, HILF M E. Identification of domains of the Tomato spotted wilt virus NSm protein involved in tubule formation, movement and symptomatology[J]. Virology, 2009, 390(1):110-121. doi: 10.1016/j.virol.2009.04.027
    [13]
    KIKKERT M, VERSCHOOR A, KORMELINK R, et al. Tomato Spotted Wilt Virus Glycoproteins Exhibit Trafficking and Localization Signals That Are FuNCtional, in Mammalian Cells[J]. Journal of Virology, 2001, 75(2):1004-1012. http://d.old.wanfangdata.com.cn/OAPaper/oai_pubmedcentral.nih.gov_113997
    [14]
    SIN S H, MCNULTY B C, KENNEDY G G, et al. Viral genetic determinants for thrips transmission of Tomato spotted wilt virus[J]. Proceedings of the National Academy of ScieNCes of the United States of America, 2005, 102(14):5168. doi: 10.1073/pnas.0407354102
    [15]
    TAKEDA A, SUGIYAMA K, NAGANO H, et al. Identification of a novel RNA sileNCing suppressor, NSs protein of Tomato spotted wilt virus[J]. Febs Letters, 2002, 532(1-2):75. doi: 10.1016-S0014-5793(02)03632-3/
    [16]
    BUCHER E, SIJEN T, HAAN P D, et al. Negative-Strand Tospoviruses and Tenuiviruses Carry a Gene for a Suppressor of Gene SileNCing at Analogous Genomic Positions[J]. Journal of Virology, 2003, 77(2):1329. doi: 10.1128-JVI.77.2.1329-1336.2003/
    [17]
    DE H P, WAGEMAKERS L, PETERS D, et al. The S RNA segment of tomato spotted wilt virus has an ambisense character[J]. Journal of General Virology, 1990, 71(5):1001-1007. doi: 10.1099/0022-1317-71-5-1001
    [18]
    KAINZ M, HILSON P, SWEENEY L, et al. Interaction between Tomato spotted wilt virus N protein monomers involves nonelectrostatic forces governed by multiple distiNCt regions in the primary structure[J]. Phytopathology, 2004, 94(7):759-65. doi: 10.1094/PHYTO.2004.94.7.759
    [19]
    RICHMOND K E, CHENAULT K, SHERWOOD J L, et al. Characterization of the Nucleic Acid Binding Properties of Tomato Spotted Wilt Virus Nucleocapsid Protein[J]. Virology, 1998, 248(1):6-11. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=9359c254d3c0bb2aac59e067cc3d19f6
    [20]
    RIBEIRO D, BORST J W, GOLDBACH R, et al. Tomato spotted wilt virus nucleocapsid protein interacts with both viral glycoproteins Gn and Gc in planta[J]. Virology, 2009, 383(1):121. doi: 10.1016/j.virol.2008.09.028
    [21]
    UHRIG J F, BUCHER G L, SCHREIER P H. The Movement Protein NSm of Tomato Spotted Wilt Tospovirus (TSWV):RNA Binding, Interaction with the TSWV N Protein, and Identification of Interacting Plant Proteins[J]. Proceedings of the National Academy of ScieNCes of the United States of America, 2000, 97(5):2373-2378. doi: 10.1073/pnas.030548397
    [22]
    JOHN A L. TRBO:A High-Efficiency Tobacco Mosaic Virus RNA-Based over-expression Vector[J]. Plant Physiology, 2007(4):1232-1240. http://d.old.wanfangdata.com.cn/NSTLQK/NSTL_QKJJ02703031/
    [23]
    孙淼, 荆陈沉, 楚成茹, 等.重庆辣椒上番茄斑萎病毒的血清学检测及分子鉴定[J].园艺学报, 2017, 44(3):487-494. http://d.old.wanfangdata.com.cn/Periodical/yyxb201703008
    [24]
    于翠, 邓凤林, 杨翠云, 等.番茄斑萎病毒外壳蛋白原核表达及Dot-blot ELISA检测方法的建立[J].浙江大学学报, 2008, 34(6):597-601. http://d.old.wanfangdata.com.cn/Periodical/zjdxxb-nyysm200806003
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(5)  / Tables(1)

    Article Metrics

    Article views (933) PDF downloads(16) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return