Monoclonal Antibodies against the Recombinant Nucleocapsid Protein of Tomato spotted wilt virus and its Application in Virus Detection
Monoclonal Antibodies against the Recombinant Nucleocapsid Protein of Tomato spotted wilt virus and its Application in Virus Detection
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DOI：
10.1111/j.1439-0434.2008.01498.x
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发表时间：
2009-06
影响因子：
1.5
通讯作者：
Jian-xiang Wu;Cui Yu;Cuiyun Yang;Xue-ping Zhou
中科院分区：
文献类型：
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作者：
Jian-xiang Wu;Cui Yu;Cuiyun Yang;Xue-ping Zhou
番茄斑枯病病毒(TSWV)是TospoVirus属的典型成员，在世界范围内对多种重要的观赏和蔬菜作物造成重大损失。以pET-32a为载体，在大肠杆菌中表达了位于猪肾综合征出血热病毒S核衣壳蛋白片段的核衣壳基因，并用抗猪肾综合征出血热病毒的单抗进行了Western印迹分析，证实重组蛋白的表达是正确的。用Ni-NTA琼脂糖凝胶对重组蛋白进行纯化，并用于生产单抗。制备了3株抗重组核衣壳蛋白的鼠源单抗。建立了三抗夹心酶联免疫吸附试验和免疫捕获RT-PCR方法，用于TSWV的可靠、高效检测。
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.