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Purification of Rice Stripe Virus.

Bio-protocol (2021-03-05)
Gang Lu, Min Yao, Yijun Zhou, Xiaorong Tao
ABSTRACT

Although many spherical and rod-shaped plant virus purification protocols are now available, only a few protocols on filamentous plant virus purification have been published. Here, we report a protocol for large-scale purification of Rice stripe virus (RSV) from RSV-infected rice tissues. RSV virions with high infectivity were first precipitated with polyethylene glycol (PEG) followed by pelleting through primary ultracentrifugation, ultracentrifugation in a glycerol cushion and ultracentrifugation in density gradient. The purified RSV virions can not only be viewed as filamentous particles under an electron microscope, but can also be acquired by insect vector through direct injection into insect body or through membrane feeding prior to transmission to rice plants.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Coomassie Brilliant blue G 250 (C.I. 42655), for electrophoresis
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
Glycerol, for molecular biology, ≥99.0%