SARS冠状病毒BJ01株基因组全长cDNA的构建与鉴定

Assembly and identification of a full-length cDNA of SARS coronavirus BJ01 strain

目的:构建我国自行分离的SARS冠状病毒BJ01株基因组全长cDNA分子,为阐明SARS病毒致病的分子机制奠定基础。方法:采用分段克隆的方法对病毒全基因组进行分段扩增和克隆,然后将全基因组各cDNA片段分别进行体外连接获得全长cDNA分子,并对其接头序列进行PCR扩增和测序鉴定。结果与结论:获得了SARS病毒BJ01株基因组全长cDNA分子,对其接头处序列的测定结果表明,该全长cDNA分子为SARS病毒BJ01株的特异序列。

Objective: To obtain the genomic full-length cDNA of SARS-CoV BJ01 strain isolated in China, and lay the foundation for clarification of the molecular mechanism of pathogenesis. Methods: First,seven cDNA fragments spanning the entire genome of SARS coronavirus were amplified by RT-PCR and cloned into vectors. Then, the subclones were digested with the appropriate restriction enzymes and assembled systematically in vitro.The junctions of cDNA were sequenced. Results and Conclusion: A full-length cDNA of SARS coronavirus was obtained and proved to be consistent with that of SARS-CoV BJ01 strain.