Identification of the epitopes of monoclonal antibodies against P74 of Helicoverpa armigera nucleopolyhedrovirus
- First Online:
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.
KeywordsHearNPV P74 Linear epitope Monoclonal antibody
Unable to display preview. Download preview PDF.
- Abbas A K, Lichtman A H, and Pillai S. 2012. Cellular and molecular immunology, Seventh edition. 2012. Elsevier Inc, p: 101. ISBN 978-1-4377-1528-6.Google Scholar
- Deng F, Wang R R, Fang M G, Jiang Y, Xu X S, Wang H Z, Chen X W, Arif B M, Guo L, Wang H L, and Hu Z H. 2007. Proteomics analysis of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus identified two new occlusion-derived virus-associated proteins, HA44 and HA100. J Virol, 81: 9377–9385.PubMedCentralPubMedCrossRefGoogle Scholar
- Haas-Stapleton E J, Washburn J O, and Volkman L E. 2004. P74 mediates specific binding of Autographa californica M nucleopolyhedrovirus occlusion-derived virus to primary cellular targets in the midgut epithelia of Heliothis virescens larvae. J Virol, 78: 6786–6791.PubMedCentralPubMedCrossRefGoogle Scholar
- Harrison R L, Sparks W O, and Bonning B C. 2010. Autographa californica multiple nucleopolyhedrovirus ODV-E56 envelope protein is required for oral infectivity and can be substituted functionally by Rachiplusia ou multiple nucleopolyhedrovirus ODV-E56. J Gen Virol, 91: 1173–1182.PubMedCrossRefGoogle Scholar
- Nie Y C, Fang M G, Erlandson M A, and Theilmann D A. 2012. Analysis of the Autographa californica multiple nucleopolyhedrovirus overlapping gene pair lef3 and ac68 reveals that AC68 is a per os infectivity factor and that LEF3 Is Critical, but not essential, for virus replication. J Virol, 86: 3985–3994.PubMedCentralPubMedCrossRefGoogle Scholar
- Ohkawa T, Washburn J O, Sitapara R, Sid E, and Volkman L E. 2005. Specific binding of Autographa californica M nucleopolyhedrovirus occlusion-derived virus to midgut cells of Heliothis virescens larvae is mediated by products of pif genes Ac119 and Ac022 but not by Ac115. J Virol, 79: 15258–15264.PubMedCentralPubMedCrossRefGoogle Scholar