期刊论文

中文

中国人兽共患病学报

1002-2694

2005

21

3

218-220,224

10.3969/j.issn.1002-2694.2005.03.007

湖南省教育厅科研项目

本校为第一机构

目的克隆SARS冠状病毒(SARS-CoV)S1基因片段,构建原核表达载体,并在宿主菌中诱导表达带组氨酸标记的融合蛋白.方法人工合成SARS-CoV S1基因片段,克隆至pUCm-T载体,经DNA序列分析和酶切后,将酶切片段亚克隆至原核表达载体pET-28b(+),重组子pET-28b/SARS-S1转化大肠杆菌ril,IPTG诱导表达,Western blot鉴定表达产物;纯化表达产物并用ELISA检测其抗原特异性.结果获得了主要以包涵体形式存在的32kD的目的蛋白,Western blot 鉴定其为带组氨酸标记的融合蛋白,ELISA证明其能与合成肽的抗体及病人血清中的SARS-CoV的抗体特异结合.结论成功构建了SARS-CoV S1基因的组氨酸标记表达载体pET-28b/ SARS-S1,并在...

To clone S1 gene fragment of SARS coronavirus (SARS-CoV) and construct the prokaryotic expression vector expressing S1 fusion protein with a 6×His-tag; S1 gene fragment is synthesized and cloned to pUCm-T vector. After sequencing , the S1 gene fragment was digested with BamH I and Sal I and inserted into prokaryotic expression vector pET-28b(+).The recombinant pET-28b(+)/ SARS-S1 was transferred into an expression strain E.coli ril and induced by IPTG to express fusion protein.The protein was identified by Western blot and purified with a kit ...