摘要
目的 :获得SARS冠状病毒S蛋白S1片段基因的克隆。方法 :将SARS病毒RNA基因组逆转录合成cDNA ,PCR得到阳性条带 ,将其亚克隆到pUCm T载体中进行酶切及测序分析 ,并与Genbank中核苷酸序列进行同源性分析。结果 :阳性克隆经酶切鉴定 ,目的基因片段长度为 1 90 5bp ,与Genbank中另外 4 5株的SARS病毒株的S蛋白S1片段基因比较 ,共有 1 3个位点发生突变 ,其中有 8处为同义突变 ,5处为错义突变。结论 :成功克隆SARS冠状病毒S蛋白S1片段基因 ,为该基因的表达及功能研究奠定了基础。
Objective:To sequence and analyze the cloned S1 fragment of spike glycoprotein gene of the SARS coronavirus.Methods:The S1 fragment of spike glycoprotein gene was obtained by RT-PCR from the SARS coronavirus genome. PCR product was ligated into pUCm-T vector and sequenced.Results:The cloned segment was 1905 bp in length. Compared with the other 45 SARS genomes available in Genbank database, 13 nucleotide variations were identified, of which 8 loci belonged to same sense mutation and 5 loci belonged to missense mutation.Conclusion:The S1 fragment of spike glycoprotein gene of the SARS coronavirus is cloned using RT-PCR,which can be used for expression and function research.
出处
《温州医学院学报》
CAS
2005年第1期16-18,共3页
Journal of Wenzhou Medical College
基金
温州市科技局基金资助项目 (Y2 0 0 3A0 0 5 )。
