Improved rapid amplification of cDNA ends (RACE) for mapping both the 5′ and 3′ terminal sequences of paramyxovirus genomes
Improved rapid amplification of cDNA ends (RACE) for mapping both the 5′ and 3′ terminal sequences of paramyxovirus genomes
复制标题
DOI：
10.1016/j.jviromet.2005.06.022
复制
发表时间：
2005-12-01
影响因子：
3.1
通讯作者：
Wang, LF
中科院分区：
文献类型：
--
作者：
Li, Z;Yu, M;Wang, LF
cDNA末端快速扩增（RACE）是一种基于PCR的RNA末端序列测定技术。然而，文献中报道的大多数RACE方法是专门针对具有3' poly-A尾和5'帽结构的真核转录物的作图而开发的。在这项研究中，开发了一种改进的RACE策略，该策略允许使用同一套常见的分子生物学试剂对副粘病毒基因组RNA进行5'和3' RACE，而不必依赖昂贵的RACE试剂盒。RNA基因组末端序列的定位是表征新型副粘病毒的重要组成部分，因为这些序列包含基因组复制和转录的重要信号，并且是研究病毒进化的重要分子标记。最近从人肾原代细胞中分离的一种新型副粘病毒的基因组两端的快速表征证明了这种策略的有用性。本文所描述的RACE策略简单、成本效益高，可用于绘制任何RNA病毒的基因组末端。(c)2005 Elsevier B. V.保留所有权利。
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. (c) 2005 Elsevier B.V. All rights reserved.