Skip to main content
ARS Home » Pacific West Area » Riverside, California » National Clonal Germplasm Repository for Citrus » Research » Publications at this Location » Publication #201435
Title: Bacterial Expression and Purification of a Protein From Citrus Leprosis Virus (CiLV)

Author
item RANGEL, E. - INIA, CENIAP, MARACAY, VE
item Keremane, Manjunath
item GUERRA-MORENO, A. - UNIV OF FL, LAKE ALFRED
item BRLANSKY, R. - UNIV OF FL, LAKE ALFRED
item Lee, Richard

Submitted to: International Organization of Citrus Virologists Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 9/1/2006
Publication Date: 1/10/2007
Citation: Rangel, E., Keremane, M.L., Guerra-Moreno, A.S., Brlansky, R.H., Lee, R.F. 2007. Bacterial Expression and Purification of a Protein From Citrus Leprosis Virus (CiLV). International Organization of Citrus Virologists Proceedings, Pg. 510.

Interpretive Summary: Citrus leprosis is a serious disease in several countries in South America and the Caribbean basin. The disease is transmitted by Brevipalpus mites. At present, the identification of the disease is based mainly on visual symptoms followed by confirmation of presence of virus particles using transmission electron microscopy. This abstract describes the expression of a protein from Citrus leprosis virus (CiLV) in a bacterial host which may be useful for development of serological assays for detection of CiLV.

Technical Abstract: Citrus leprosis is a serious disease in several countries in South America and the Caribbean basin. The disease is transmitted by Brevipalpus mites. At present, the identification of the disease is based mainly on visual symptoms followed by confirmation of presence of virus particles using transmission electron microscopy. Early identification of the disease from plant materials and mites is important for management of the disease and to minimize spread into new areas. A highly expressed protein from the putative cytoplasmic leprosis virus genome was cloned in a bacterial expression vector, pET 27b (Novagen). The leprosis protein with a carboxy-terminal fusion of a HSV and 6x Histidine tags was expressed in the expression host, Escherichia coli, strain BL21. The expressed protein was purified by two methods; the insoluble inclusion protein was purified by using Bugbuster (Novagen), and the soluble fraction protein was purified using a Ni-NTA agarose column. The expressed protein was monitored through different steps of purification by Western blotting using an anti-HSV monoclonal antibody. Both the expressed protein preparations were used as inject antigens in rabbits and chickens for producing antibodies to CiLV.