A simple method for cloning the complete begomovirus genome using the bacteriophage φ29 DNA polymerase

被引：296

作者：

Inoue-Nagata, AK

Albuquerque, LC

Rocha, WB

Nagata, T

机构：

[1] Embrapa Hortalicas, BR-70359970 Brasilia, DF, Brazil

[2] Univ Catolica Brasilia, BR-70790160 Brasilia, DF, Brazil

来源：

JOURNAL OF VIROLOGICAL METHODS | 2004年 / 116卷 / 02期

关键词：

circular DNA; Geminiviridae; rolling circle;

D O I：

10.1016/j.jviromet.2003.11.015

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 2003 Elsevier B.V. All rights reserved.

引用

页码：209 / 211

页数：3

共 10 条

[1]

[Anonymous], 2000, VIRUS TAXONOMY 7 REP

[2]

Dellaporta S. L., 1983, Plant Molecular Biology Reporter, V1, P19, DOI [10.1007/BF02712670, DOI 10.1007/BF02712670]

[3] CLONING OF THE COMPLETE DNA GENOMES OF 4 BEAN-INFECTING GEMINIVIRUSES AND DETERMINING THEIR INFECTIVITY BY ELECTRIC-DISCHARGE PARTICLE-ACCELERATION [J].