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J Virol Methods. 2015 Dec 1;225:23-9. doi: 10.1016/j.jviromet.2015.08.023. Epub 2015 Sep 2.

Highly efficient strategy for the heterologous expression and purification of soluble Cowpea chlorotic mottle virus capsid protein and in vitro pH-dependent assembly of virus-like particles.

Author information

1
Biomolecular Diversity Laboratory, Unidad de Genómica Avanzada, Centro de Investigación y Estudios Avanzados del Instituto Politécnico Nacional, Unidad Irapuato, Mexico.
2
Laboratorio de Interacciones Planta-Virus at Departamento de Ingeniería Genética, Centro de Investigación y Estudios Avanzados del Instituto Politécnico Nacional, Unidad Irapuato, Mexico.
3
Biomolecular Diversity Laboratory, Unidad de Genómica Avanzada, Centro de Investigación y Estudios Avanzados del Instituto Politécnico Nacional, Unidad Irapuato, Mexico. Electronic address: trippm@langebio.cinvestav.mx.

Abstract

Obtaining pure and soluble viral capsid proteins (CPs) has been a major challenge in the fields of science and technology in recent decades. In many cases, the CPs can self-assemble in the absence of a viral genome, resulting in non-infectious, empty virus-like particles (VLPs) which can be safely handled. The use of VLPs has found great potential in biotechnology and health purposes. In addition, VLPs are a good model system to study protein-protein interactions at the molecular level. In this work, an optimized strategy for the heterologous expression of the Cowpea chlorotic mottle virus (CCMV) CP based in Escherichia coli is described. The method is efficient, inexpensive and it consistently produces higher yields and greater purity levels than those reported so far. Additionally, one of the main advantages of this method is the prevention of the formation of inclusion bodies, thus allowing to directly obtain high amounts of the CP in a soluble and functionally active state with the capacity to readily form VLPs in vitro. The CCMV CP self-assembly pH dependence was also investigated, providing guidelines to easily modulate the process.

KEYWORDS:

Capsid protein; Cowpea chlorotic mottle virus (CCMV); In vitro virus assembly; Protein expression purification; Virus like particles

PMID:
26342905
DOI:
10.1016/j.jviromet.2015.08.023
[Indexed for MEDLINE]

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