Identification of the Epitopes of Monoclonal Antibodies against P74 of <i>Helicoverpa armigera</i> Nucleopolyhedrovirus

Limin Liao; Dianhai Hou; Huachao Huang; Manli Wang; Fei Deng; Hualin Wang; Zhihong Hu; Tao Zhang

doi:10.1007/s12250-013-3393-7

December 2013

Limin Liao, Dianhai Hou, Huachao Huang, Manli Wang, Fei Deng, Hualin Wang, Zhihong Hu and Tao Zhang. Identification of the Epitopes of Monoclonal Antibodies against P74 of Helicoverpa armigera Nucleopolyhedrovirus[J]. Virologica Sinica, 2013, 28(6): 360-367. doi: 10.1007/s12250-013-3393-7

Citation: Limin Liao, Dianhai Hou, Huachao Huang, Manli Wang, Fei Deng, Hualin Wang, Zhihong Hu, Tao Zhang. Identification of the Epitopes of Monoclonal Antibodies against P74 of Helicoverpa armigera Nucleopolyhedrovirus .VIROLOGICA SINICA, 2013, 28(6) : 360-367. http://dx.doi.org/10.1007/s12250-013-3393-7

Identification of the Epitopes of Monoclonal Antibodies against P74 of Helicoverpa armigera Nucleopolyhedrovirus

cstr: 32224.14.s12250-013-3393-7

Skate Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

通讯作者： Tao Zhang, tzhang@wh.iov.cn
收稿日期： 2013-10-11
录用日期： 2013-11-25
出版日期： 2013-12-02

摘要

P74 is a per os infectivity factor of baculovirus. Here, we report the production of three monoclonal antibodies (mAbs), denoted as 20D9, 20F9 and 21E1, raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV), and the identification of their recognition epitopes. The full-length P74, without the transmembrane domains at the C-terminus, was first divided into three segments (N, M and C, respectively), based on the proposed cleavage model for the protein, which were then expressed individually. Western blot analyses revealed specific cross-reactions with the N fragment, for both 20D9 and 21E1. Extensive truncation, followed by prokaryotic expression, of the P74 N fragment was then performed in order to screen for linear epitopes of P74. The recognition regions of 20D9 and 21E1 were revealed to be localized at R144-T153 and T199-C219, respectively. In addition, immunofluorescence microscopy indicated that 20D9 and 20F9 could recognize native P74 in HearNPV-infected cells. These findings will facilitate further investigations of the proteolytic processing of HearNPV P74, and of its involvement in virus-host interactions.
- HearNPV
- , P74
- , Linear epitope
- , Monoclonal antibody

Identification of the Epitopes of Monoclonal Antibodies against P74 of Helicoverpa armigera Nucleopolyhedrovirus

Skate Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

Corresponding author: Tao Zhang, tzhang@wh.iov.cn
Received Date: 11 October 2013
Accepted Date: 25 November 2013
Published Date: 02 December 2013

Abstract

P74 is a per os infectivity factor of baculovirus. Here, we report the production of three monoclonal antibodies (mAbs), denoted as 20D9, 20F9 and 21E1, raised against P74 of Helicoverpa armigera nucleopolyhedrovirus (HearNPV), and the identification of their recognition epitopes. The full-length P74, without the transmembrane domains at the C-terminus, was first divided into three segments (N, M and C, respectively), based on the proposed cleavage model for the protein, which were then expressed individually. Western blot analyses revealed specific cross-reactions with the N fragment, for both 20D9 and 21E1. Extensive truncation, followed by prokaryotic expression, of the P74 N fragment was then performed in order to screen for linear epitopes of P74. The recognition regions of 20D9 and 21E1 were revealed to be localized at R144-T153 and T199-C219, respectively. In addition, immunofluorescence microscopy indicated that 20D9 and 20F9 could recognize native P74 in HearNPV-infected cells. These findings will facilitate further investigations of the proteolytic processing of HearNPV P74, and of its involvement in virus-host interactions.
- HearNPV
- , P74
- , Linear epitope
- , Monoclonal antibody

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