• 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 38 Issue 3
Mar.  2023
Turn off MathJax
Article Contents
CHEN C, GAO Y F, WANG X R, et al. Preparation of Polyclonal Antibody against MiPDCD6 Protein in Meloidogyne incognita [J]. Fujian Journal of Agricultural Sciences,2023,38(3):322−328 doi: 10.19303/j.issn.1008-0384.2023.03.008
Citation: CHEN C, GAO Y F, WANG X R, et al. Preparation of Polyclonal Antibody against MiPDCD6 Protein in Meloidogyne incognita [J]. Fujian Journal of Agricultural Sciences,2023,38(3):322−328 doi: 10.19303/j.issn.1008-0384.2023.03.008

Preparation of Polyclonal Antibody against MiPDCD6 Protein in Meloidogyne incognita

doi: 10.19303/j.issn.1008-0384.2023.03.008
  • Received Date: 2022-10-09
  • Rev Recd Date: 2022-12-22
  • Available Online: 2023-03-28
  • Publish Date: 2023-03-28
  •   Objective   The polyclonal antibody against MiPDCD6 protein in the esophageal glands of Meloidogyne incognita was prepared to study the pathogenic mechanism of the root-knot disease on plants transmitted by the nematode.   Method  The functional fragment of MiPDCD6 was amplified by PCR to construct recombinant plasmid pET-32a-MiPDCD6 and transform it into Escherichia coli BL21 cells for MiPDCD6 fusion protein induction and expression. Polyclonal antibodies were prepared by immunizing male New Zealand white rabbits with purified MiPDCD6 expression protein. Titer and purification of the obtained antibody were verified using ELISA and SDS-PAGE techniques.   Result  Under IPTG concentration of 1.0 mmol·L−1 at 37℃with constant rotation of 150 r·min−1 for 5 h, MiPDCD6 transformed in the E. coli BL21 was clearly expressed. The secured polyclonal antibody was highly specific with a high titer of approximately 1∶50000 as shown by ELISA and SDS-PAGE.   Conclusion   The prokaryotic expression conditions of MiPDCD6 were determined. The polyclonal antibody obtained had a high titer and specification against MiPDCD6 and was considered adequate for studying the pathogenesis of the root knot disease on plants infected through M. incognita.
  • loading
  • [1]
    JONES J T, HAEGEMAN A, DANCHIN E G J, et al. Top 10 plant-parasitic nematodes in molecular plant pathology [J]. Molecular Plant Pathology, 2013, 14(9): 946−961. doi: 10.1111/mpp.12057
    [2]
    MAKI M, SUZUKI H, SHIBATA H. Structure and function of ALG-2, a penta-EF-hand calcium-dependent adaptor protein [J]. Science China Life Sciences, 2011, 54(8): 770−779. doi: 10.1007/s11427-011-4204-8
    [3]
    MAKI M, TAKAHARA T, SHIBATA H. Multifaceted roles of ALG-2 in Ca2+-regulated membrane trafficking [J]. International Journal of Molecular Sciences, 2016, 17(9): 1401. doi: 10.3390/ijms17091401
    [4]
    HENZL M T. Ligation events influence ALG-2 dimerization [J]. Biophysical Chemistry, 2018, 239: 16−28. doi: 10.1016/j.bpc.2018.05.001
    [5]
    RAO R V, POKSAY K S, CASTRO-OBREGON S, et al. Molecular components of a cell death pathway activated by endoplasmic reticulum stress [J]. Journal of Biological Chemistry, 2004, 279(1): 177−187. doi: 10.1074/jbc.M304490200
    [6]
    HUANG G, ALLEN R, DAVIS E L, et al. Engineering broad root-knot resistance in transgenic plants by RNAi silencing of a conserved and essential root-knot nematode parasitism gene [J]. Proceedings of the National Academy of Sciences, 2006, 103(39): 14302−14306. doi: 10.1073/pnas.0604698103
    [7]
    谢家廉. 南方根结线虫效应蛋白Misp12功能分析和淡紫紫孢菌寄生适应性的比较基因组研究[D]. 武汉: 华中农业大学, 2016.

    XIE J L. Functional analysis study of the Meloidogyne incognita effector Misp12/genomic and transcriptom study of the nematophagous Purpureocillium lilacinum 36-1 [D]. Wuhan: Huazhong Agricultural University, 2016. (in Chinese)
    [8]
    KAMARUZZAMAN M, ZHAO L F, ZHANG J A, et al. MiPDCD6 effector suppresses host PAMP-triggered immunity to facilitate Meloidogyne incognita parasitism in tomato [J]. Plant Pathology, 2022, 72(1): 195−206.
    [9]
    吴路平. 南方根结线虫MiPDCD6蛋白抑制寄主免疫的相关基因克隆及表达分析[D]. 广州: 华南农业大学, 2018

    WU L P. Express analysis of MiPDCD6, LeMYB330 and cloning LeCyt-C involved in the process of root-knot nematode(Meloidogyne incognita) to suppress plant immunity in tomato(Lycopersicon esculentum)[D]. Guangzhou: South China Agricultural University, 2018. (in Chinese)
    [10]
    贾慧娜, 罗海玲. 多克隆抗体制备方法的研究进展 [J]. 中国草食动物科学, 2012, 32(S1):66−69. doi: 10.3969/j.issn.2095-3887.2012.z1.022

    JIA H N, LUO H L. Progress in preparation of polyclonal antibodies [J]. China Herbivore Science, 2012, 32(S1): 66−69.(in Chinese) doi: 10.3969/j.issn.2095-3887.2012.z1.022
    [11]
    NEWCOMBE A R, CRESSWELL C, Davies S, et al. Optimised affinity purification of polyclonal antibodies from hyper immunized ovine serum using a synthetic: Protein A adsorbent, MAbsorbent A2P [J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2005, 814(2): 209−215. doi: 10.1016/j.jchromb.2004.10.027
    [12]
    UMA M, RAO P P, NAGALEKSHMI K, et al. Expression and purification of polioviral proteins in E. coli, and production of antisera as reagents for immunological assays [J]. Protein expression and purification, 2016, 128: 115−122. doi: 10.1016/j.pep.2016.08.014
    [13]
    SAHDEV S, KHATTAR S K, SAINI K S. Production of active eukaryotic proteins through bacterial expression systems: A review of the existing biotechnology strategies (Review) [J]. Molecular and Cellular Biochemistry, 2008, 307: 249−264.
    [14]
    仇书兴, 殷星, 苏淑娟, 等. 新型H7N9禽流感病毒NA蛋白胞外区片段的生物信息学分析及其多克隆抗体制备 [J]. 生物技术通报, 2019(12):85−93.

    QIU S X, YIN X, SU S J, et al. Bioinformatics analysis of the extracellular region of NA protein in novel H7N9 avian influenza virus and preparation of polyclonal antibodies [J]. Biotechnology Bulletin, 2019(12): 85−93.(in Chinese)
    [15]
    苗碧琛, 陈松彪, 张秀娟, 等. 猪细小病毒NS基因原核表达与多克隆抗体制备 [J]. 动物医学进展, 2020, 41(2):1−8. doi: 10.3969/j.issn.1007-5038.2020.02.001

    MIAO B C, CHEN S B, ZHANG X J, et al. Prokaryotic expression of Porcine parvovirus NS gene and preparation of polyclonal antibody [J]. Progress in Veterinary Medicine, 2020, 41(2): 1−8.(in Chinese) doi: 10.3969/j.issn.1007-5038.2020.02.001
  • 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(4)

    Article Metrics

    Article views (572) PDF downloads(11) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return