我国SARS病毒BJ01株基因组亚cDNA克隆的构建与鉴定

目的：构建我国SAILS病毒BJ01株基因组全序列的cDNA克隆，为进一步构建该毒株的全长感染性cDNA克隆奠定基础。方法：根据BJ01株病毒基因组序列中含有的特异酶切位点，利用DNAstar软件设计7对引物。然后通过RT-PCR扩增出覆盖病毒基因组全长的7个cDNA片段，并分别将其克隆至pGEM-T Easy或Topo载体。结果与结论：已构建出SARS-CoV BJ01株基因组的7个亚cDNA克隆，经酶切鉴定和序列测定表明，所获得的cDNA克隆为BJ01株病毒的特异序列。

Construction and identification of genomic cDNA subclones of SARS-CoV BJ01 strain in China

Objective: To get the cDNA subclones spanning the entire genome of SARS coronavirus BJ01 strain, and lay the foundation for construction of genomic full-length infectious cDNA clone. Methods: According to the restriction endonuclease sites in viral genome of BJ01 strain, seven primer pairs were designed by DNAstar software and the cDNA fragments were amplified by RT-PCR.The amplified products were purified and then cloned into pGEM-T Easy or Topo vectors. Results and Conclusions: Seven cDNA subclones covering the viral genome were obtained. The subclones were sequenced and proved to be consistent with SARS-CoV BJ01 strain.