English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

A large scale purification is required for biochemical and biophysical research of aphid-borne circulative plant viruses, because of the low virus content in the host plants. The current procedure for purification of potato leaf-roll virus (PLRV) from plant materials is fairly laborious1). Since Hebert2) reported polyethylene glycol (PEG) precipitation of plant viruses, many workers have applied this PEG precipitation for purifications of bacteriophages and animal viruses3,4). We have applied PEG precipitation to PLRV, as a modification of the purification procedure described previously1). In preliminary experiments, assays by aphid-injection showed that precipitates obtained by the addition of 4 or 8% PEG to clarified plant juice retained infectivity. However, the amount of virus in the precipitates could not be accurately measured by the bioassay. Accordingly, comparative assays using precipitates obtained by the addition of PEG at various concentrations were made by an analytical sucrose density-gradient centrifugation5). Frozen tissue of Physalis floridana plants infected with PLRV was ground by an electric meat grinder. The macerate was then homogenized with a Waring Blendor in 0.5M phosphate buffer (pH 7.4). The homogenate was emulsified with a chloroform and n-butanol mixture (1:1) in a fruit mixer. The preparation was clarified by centrifuging at 6,000rpm for 15 minutes in 9B-2 rotor of a Marusan high-speed centrifuge. PEG 6,000 was added at various concentrations to the resultant aqueous phase, which was stirred for 60 minutes and then kept in a centrifuge tube for 60 minutes at room temperature. Precipitates obtained after centrifugation in the same rotor at 6,000rpm for 15 minutes were suspended in 0.01M phosphate buffer (pH 7.4) with a chilled Teflon-glass homogenizes. The resuspended preparation was again clarified with fluorocarbon (Daifron S-3) in a cold room, and further purified by two cycles of differential centrifugation (at 40,000rpm for 120 minutes; at 10,000rpm for 15 minutes) in RP40 rotor of a refrigerated Hitachi 40P ultracentrifuge. Two milliliters each of the concentrated preparation (from 400g of tissue) was layered on a sucrose gradient column, and was centrifuged at 23,000rpm for 180 minutes in RPS25 rotor of the same ultracentrifuge. Density-gradient columns, made by layering of 4, 7, 7 and 7ml of 100, 200, 300 and 400mg of sucrose

The Application of Polyethylene Glycol Precipitation for Purification of Potato Leaf-Roll Virus