Monoclonal Antibodies Against the Whitefly-Transmitted Tomato Yellow Leaf Curl Virus and Their Application in Virus Detection

WU Jian-xiang; SHANG Hai-li; XIE Yan; SHEN Qing-tang; ZHOU Xue-ping

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Monoclonal Antibodies Against the Whitefly-Transmitted Tomato Yellow Leaf Curl Virus and Their Application in Virus Detection

机译：抗粉虱传播的番茄黄叶卷毛病毒单克隆抗体及其在病毒检测中的应用

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Tomato yellow leaf curl virus(TYLCV)is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein(CP)gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21(DE3)using pET-32a as the expression vector.The recombinant protein was purified through Ni+-NTA affinity column and used to immunize BALB/c mice.Three hybridoma cell lines(2B2,2E3 and 3E10)secreting monoclonal antibodies(MAbs)against TYLCV CP were obtained by fusing mouse myeloma cells(Sp 2/0)with spleen cells from the immunized BALB/c mouse.The titers of ascitic fluids of three MAbs ranged from 10-6 to 10-7 in indirect-ELISA.Isotypes and subclasses of all the MAbs belonged to IgG1,κ light chain.Triple antibody sandwich enzyme-linked immunosorbent assay(TAS-ELISA)showed that the MAb 3E10 could react with five begomoviruses infecting tomato,while the other two(2B2 and 2E3)mainly reacted with TYLCV.TAS-ELISA was set up using the MAb 3E10,and the established method could successfully detect virus in plant sap at 1:2 560(w/v,g mL-1).Detection of field samples showed that begomoviruses were common in tomato crops in Zhejiang Province,China.

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《农业科学学报（英文版）》 |2012年第2期|263-268|共6页
作者
WU Jian-xiang; SHANG Hai-li; XIE Yan; SHEN Qing-tang; ZHOU Xue-ping;
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Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China;

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China;

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China;

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China;

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China;

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Monoclonal Antibodies Against the Whitefly-Transmitted Tomato Yellow Leaf Curl Virus and Their Application in Virus Detection

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