Rolling circle amplification revolutionizes diagnosis and genomics of geminiviruses

被引：319

作者：

Haible, Daniela ^{[1
]}

Kober, Sigrid ^{[1
]}

Jeske, Holger ^{[1
]}

机构：

[1] Univ Stuttgart, Dept Mol Biol & Plant Virol, D-70550 Stuttgart, Germany

来源：

JOURNAL OF VIROLOGICAL METHODS | 2006年 / 135卷 / 01期

关键词：

geminivirus; diagnosis; sequencing; infectivity;

D O I：

10.1016/j.jviromet.2006.01.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Better, easier and cheaper than polymerase chain reaction (PCR) or antibody detection, rolling circle amplification (RCA) using the bacteriophage 29 DNA polymerase allows for a reliable diagnosis of geminiviruses and presumably all viruses with small single-stranded circular DNA genomes. The results show the efficiency of this technique in characterizing viral DNA components of several geminiviruses from experimental and natural host plant sources. The advantages are: (a) that no expensive devices are necessary, (b) simple handling, (c) detection of all infecting circular DNA components without any knowledge of sequence information in a single step, and (d) low costs per reaction. In addition, RCA-amplified viral DNA can be characterized by restriction fragment length polymorphism analysis and directly sequenced up to 900 bases in a single run circumventing cloning and plasmid purification. This shortcut will considerably accelerate genomics of at least gemini-, nano- and circoviruses in the future. (c) 2006 Elsevier B.V. All rights reserved.

引用

页码：9 / 16

页数：8

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