A simple method for cloning the complete begomovirus genome using the bacteriophage phi29 DNA polymerase.

Abstract:

:The bacteriophage phiDNA polymerase amplifies circular DNA in a rolling circle amplification mechanism. This characteristic was applied to amplify and clone the complete circular DNA genome of a begomovirus. Total DNA extracted from infected tissue was used as the template of an amplification reaction using the commercial kit TempliPhi (Amersham Biosciences). The amplified DNA could be used for direct sequencing and was cloned after digestion with a single cutting restriction endonuclease. The use of this enzyme simplified the cloning steps and increased the cloning efficiency of the complete genome of a circular plant DNA virus.

journal_name

J Virol Methods

authors

Inoue-Nagata AK,Albuquerque LC,Rocha WB,Nagata T

doi

10.1016/j.jviromet.2003.11.015

subject

Has Abstract

pub_date

2004-03-15 00:00:00

pages

209-11

issue

2

eissn

0166-0934

issn

1879-0984

pii

S0166093403003938

journal_volume

116

pub_type

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