Detection and sequence characterization of the 3'-end of coronavirus genomes harboring the highly conserved RNA motif s2m.
Detection and sequence characterization of the 3'-end of coronavirus genomes harboring the highly conserved RNA motif s2m.
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DOI：
10.1007/978-1-59745-181-9_3
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Jonassen CM
中科院分区：
文献类型：
--
作者：
Jonassen CM
在Astroviridae、Coronaviridae和PicorNaviridae的几个成员的基因组的3‘端存在一个非常保守的43个核苷酸长的基序，可以用于检测和分析携带它的病毒的序列。该程序利用位于S2m最保守核心的引物在半特异性PCR中朝向普通锚定寡核苷酸(DT)引物。这一策略允许从病毒基因组的3‘端对大约50-100个核苷酸进行测序，为启动快速扩增cDNA端(5’-RACE)和引物走动策略的进一步基因组特征提供了足够的序列信息。
A remarkably conserved 43-nucleotide-long motif present at the 3′-end of the genomes of several members of the polyadenylated RNA virus families Astroviridae, Coronaviridae, and Picornaviridae can be used for the detection and sequence characterization of the viruses harboring it. The procedure makes use of a primer located in the most conserved core of s2m toward a generic anchored oligo(dT) primer in a semispecific PCR. This strategy allows the sequencing of some 50–100 nucleotides from the 3′-end of the virus genome, representing sufficient sequence information for initiation of further genomic characterization in a rapid amplification of cDNA ends (5′-RACE) and primer walking strategy.