Sequence Analysis of the Terminal Region of Dengue Fever Virus Genome RNA
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摘要:
采用磁性分离技术从登革热病毒(D2-04株)感染的C6/36细胞中分离了D2-04病毒RNA.以该RNA为模板进行RT-PCR,分别扩增了D2-04 RNA 5′和3′端cDNA片段,该cDNA片段分别克隆到pGEM-3Z质粒多聚接头的HincⅡ位点得到含有5′端284 bp及3′端525 bp cDNA的重组质粒.通过荧光标记引物及双脱氧核苷酸PCR方法测定了上述cDNA插入片段的序列.同源性比较结果证明D2-04株与其他不同株间的同源性较高,可达93%～98%;不同型间的同源性较差, 仅80%左右; 属间的同源性更低.
Abstract:
D2-04 RNA was isolated magnetically from cultured C6/36 cells. With this RNA as template, the cDNA both 5′ and 3′-tumini were amplified by RT-PCR respectively. After inserted into pGEM-3Z plasmid, the cDNAs were sequenced by dye-primer and dye terminator PCR sequencing strategy, and the primary and stable secondary RNA structure were analyzed by computer software on VAX-11-780 computer.
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