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Monoclonal Antibodies against the Recombinant Nucleocapsid Protein of Tomato spotted wilt virus and its Application in Virus Detection
Author(s) -
Wu Jianxiang,
Yu Cui,
Yang Cuiyun,
Zhou Xueping
Publication year - 2009
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.2008.01498.x
Subject(s) - biology , virology , recombinant dna , tospovirus , monoclonal antibody , virus , antibody , plant virus , microbiology and biotechnology , dot blot , western blot , tomato spotted wilt virus , gene , immunology , biochemistry
Tomato spotted wilt virus (TSWV) is the type member of the tospovirus genus and causes significant losses in a wide range of economically important ornamental and vegetable crops worldwide. The nucleocapsid gene, located on the ambisense S RNA segment of TSWV was expressed in Escherichia coli using pET‐32a as vector and correct expression of recombinant protein was confirmed by Western blot using an anti‐TSWV monoclonal antibody (MAb). The recombinant protein was purified using Ni‐NTA agarose and the purified protein was used for the production of MAbs. Three murine MAbs against the recombinant nucleocapsid protein were produced. Triple antibody sandwich enzyme‐linked immunosorbent assay and immunocapture RT‐PCR methods were then established for reliable and efficient detection of TSWV using the produced MAbs.