Simple and Robust <em>in vivo</em> and <em>in vitro</em> Approach for Studying Virus Assembly

Sonali Chaturvedi; Bongsu Jung; Sharad Gupta; Bahman Anvari; A.L.N. Rao

doi:10.3791/3645

Simple and Robust in vivo and in vitro Approach for Studying Virus Assembly

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Simple and Robust in vivo and in vitro Approach for Studying Virus Assembly

Simple and Robust in vivo and in vitro Approach for Studying Virus Assembly

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09:47 min

March 1, 2012

DOI: 10.3791/3645-v

Sonali Chaturvedi¹, Bongsu Jung², Sharad Gupta², Bahman Anvari², A.L.N. Rao¹

¹Department of Plant Pathology and Microbiology,University of California, Riverside , ²Department of Bioengineering,University of California, Riverside

A simple, efficient and robust way to synchronize the delivery of multiple viral components to plant cells via Agrobacterium-mediated transient expression is described. This approach is amenable for studying replication, encapsidation followed by in vitro reassembly of non-viral components into genome depleted optical viral ghosts suitable for biomedical applications.

The overall goal of this procedure is to study virus assembly in vivo and in vitro. This is accomplished by first expressing bro mosaic virus RNAs and bacteria, and exposing it to plant cells through agro infiltration. Next, the BMV virions are purified from infiltrated leaves.

Then capsid protein subunits for in vitro assembly are prepared. Finally, BMV Varians are assembled in vitro using the capsid protein and indocyanine green dye. Ultimately, the results show in vitro assembled bro mosaic varians through formation of optical ghosts, which can be determined by absorbance and emission of fluorescence.

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