Characterisation of visna virus reverse transcriptase: a micro scale reverse transcriptase assay adapted for use with an automated cell harvester.

Abstract:

:The reverse transcriptase of the sheep lentivirus visna/maedi virus has been characterised. Optima for magnesium ion concentration (5-10 mM), potassium ion concentration (150 mM) and pH (8.25) for this enzyme are very similar to those previously described for the human immunodeficiency viruses. The assay used for this work makes use of a cell harvester to speed up the processing of multiple samples. It is small scale, requiring 15 microliters of sample, is rapid, and is able to detect virus at titres below 10(3)/ml. Harvesting the assay onto either DEAE paper or using TCA and glass fibre mats make it suitable for use with either tissue culture media or infected cell lysates, but not with body fluids. It has been used to detect cell-associated reverse transcriptase in choroid plexus cells within 36 h of visna infection.

journal_name

J Virol Methods

authors

Sargan DR,Watt NJ,McConnell I

doi

10.1016/0166-0934(92)90144-3

subject

Has Abstract

pub_date

1992-02-01 00:00:00

pages

129-40

issue

2

eissn

0166-0934

issn

1879-0984

pii

0166-0934(92)90144-3

journal_volume

36

pub_type

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