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 co-precipitants. The crystals belonged to the cubic space group P23, with unit-cell parameter a = 392 A, and diffracted X-rays to at least 4.5 A resolution.
Lee
Kian-Chung
KC
Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609, Singapore.
Lim
Daina
D
Wong
Sek-Man
SM
Dokland
Terje
T
D002072:Butylene Glycols / Q000737:chemistry; D019184:Carmovirus / Q000737:chemistry*; D002499:Centrifugation, Density Gradient; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D008854:Microscopy, Electron; D009991:Oscillometry; D018515:Plant Leaves / Q000821:virology; D011092:Polyethylene Glycols / Q000494:pharmacology; D014961:X-Ray Diffraction
D002072:Butylene Glycols; D011092:Polyethylene Glycols; C026978:2,3-butylene glycol