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A typical project workflow includes: (1) the preparation of homogenous virus samples either in eukaryotic cell cultures or amoeba cultures (for giant viruses); (2) preparation of complexes with neutralizing antibodies, anti-viral compounds or receptors; (3) followed by freezing grids for single particle cryo-EM or cryo-ET studies. Standard molecular virology assays, mutational studies and molecular modeling complement structural studies. A few of my ongoing projects (in the Kuhn and the Rossmann labs) are described below: 

(1) Structure of Zika virus and its complex with neutralizing antibodies

We aim to determine the high resolution Cryo-EM structures of Zika virus (ZIKV) and its complexes with antibodies to decipher the structural components of ZIKV infection mechanism and its neutralization. The epitopes of diverse human monoclonal antibodies against ZIKV are mapped using structural data. This is crucial for structure based design of therapeutic antibodies. (See CV/Publications: 2, 3, 4, 32, 34)

Movie: Cryo-EM structure of Zika virus at a resolution of 3.1 Å.

(2) Near atomic resolution structure of Pacmanvirus, a 250 nm dsDNA giant virus related to ASFV

Here, we aim to deduce the structure of Pacmanvirus to identify major and minor capsid proteins and understand capsid assembly in giant viruses. Single particle cryo-EM and localized 3D reconstruction methods are being used to generate high resolution maps. (See CV/Publications: 5, 33)




Figure: (A) Micrographs of Pacmanvirus, (B) electrostatic potential map after single particle 3D reconstruction and (C) icosahedral model of faustovirus (EMD-8144), a representative and related giant virus to pacmanvirus.

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