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Improved rapid amplification of cDNA ends (RACE) for mapping both the 5' and 3' terminal sequences of paramyxovirus genomes.

Abstract：

:Rapid amplification of cDNA ends (RACE) is a powerful PCR-based technique for determination of RNA terminal sequences. However, most of the RACE methods reported in the literature are developed specifically for the mapping of eukaryotic transcripts with 3' poly-A tail and 5' cap structure. In this study, an improved RACE strategy was developed which allows both 5' and 3' RACE of paramyxovirus genomic RNA using the same set of common molecular biology reagents without having to rely on expensive RACE kits. Mapping of RNA genome terminal sequences is an essential part of characterizing novel paramyxoviruses since these sequences contain important signals for genome replication and transcription, and are important molecular markers for studying virus evolution. The usefulness of this strategy was demonstrated by rapid characterization of both genome ends for a novel paramyxovirus recently isolated from human kidney primary cells. The RACE strategy described in this paper is simple, cost-effective and can be used to map genome ends of any RNA viruses.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Li Z,Yu M,Zhang H,Wang HY,Wang LF

doi

10.1016/j.jviromet.2005.06.022

subject

Has Abstract

pub_date

2005-12-01 00:00:00

pages

154-6

issue

1-2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(05)00210-7

journal_volume

130

pub_type

杂志文章

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