Purification, crystallization and X-ray analysis of Hibiscus chlorotic ringspot virus

Hibiscus chlorotic ringspot virus (HCRSV), a Carmovirus, occurs worldwide and induces chlorotic ringspots on leaves, stunting and flower distortion in Hibiscus species, including kenaf. The HCRSV capsid has T = 3 icosahedral symmetry and contains 180 copies of the coat protein. A virus yield of 48 - 70 mg per 100 g of infected kenaf leaves was achieved with an improved purification scheme involving sucrose-cushion and sucrose density-gradient centrifugation. The virus was crystallized using PEG 8000 and 2,3-butanediol as coprecipitants. The crystals belonged to the cubic space group P23, with unit-cell parameter a = 392 Angstrom, and diffracted X-rays to at least 4.5 Angstrom resolution.