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| 丁型肝炎病毒ORF5基因片段的原核表达及在ELISA中的初步应用 | |
| 引用本文： | 王云龙,王娟,李玉林,董彩文,孙新城,张怡青,王国强,李恒思,刘旺根. 丁型肝炎病毒ORF5基因片段的原核表达及在ELISA中的初步应用[J]. 动物医学进展, 2012, 33(5): 31-34 |
| 作者姓名： | 王云龙 王娟 李玉林 董彩文 孙新城 张怡青 王国强 李恒思 刘旺根 |
| 作者单位： | 1. 河南师范大学,河南新乡453007;郑州职业技术学院,河南郑州450001 2. 河南师范大学,河南新乡,453007 3. 河南省生物工程技术研究中心,河南郑州,450001 |
| 基金项目： | 河南省科技创新团队和郑州市科技创新团队项目 |
| 摘 要： | 为获得HDAg抗原,以含有ORF5编码区基因的pETHD质粒为模板进行PCR扩增得到500bp大小片段,经过BamHⅠ、HindⅢ双酶切,与载体pET-30a连接,构建重组表达质粒pET-30a/HDAg,再转入细胞BL21(DE3),经过IPTG诱导表达和SDS-PAGE分析,表达蛋白分子质量约27ku,经过Ni-NTA亲和层析纯化后,纯度达95%以上。将纯化的蛋白经HRP标记,初步建立ELISA捕获法,通过对24份标本检测和交叉反应试验,结果表明表达的抗原具有很好的反应原性和特异性,为建立丁型肝炎早期检测试剂盒提供材料。 |
| 关 键 词： | 丁型肝炎病毒 丁肝抗原 原核表达 ELISA |
Prokaryotic Expression of Hepatitis Delta Virus ORF5 Gene and Its Application in ELISA |
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| WANG Yun-long , WANG Juan , LI Yu-lin , DONG Cai-wen , SUN Xin-cheng , ZHANG Yi-qing , WANG Guo-qiang , LI Heng-si , LIU Wang-gen. Prokaryotic Expression of Hepatitis Delta Virus ORF5 Gene and Its Application in ELISA[J]. Progress In Veterinary Medicine, 2012, 33(5): 31-34 | |
| Authors: | WANG Yun-long WANG Juan LI Yu-lin DONG Cai-wen SUN Xin-cheng ZHANG Yi-qing WANG Guo-qiang LI Heng-si LIU Wang-gen |
| Affiliation: | 1.Henan Normal University,Xinxiang,Henan,453007,China;2.Zhengzhou Technical College,Zhengzhou, Henan,450001,China;3.Henan Biotechnology Research Center,Zhengzhou,Henan,450001,China) |
| Abstract: | To obtain recombinant hepatitis delta virus antigen,the fragments of HDAg amplified by PCR from pETHD was inserted into expression vector pET-30a.The recombinant plasmid was transformed into BL21(DE3),where hepatitis delta virus antigen was induced to express by IPTG.Analyzed by SDS-PAGE,the product was 27 ku.Purified by Ni-NTA metal-affinity chromatography,protein’s purity was 95%.The HRP-labeled protein was used to detecte 24 samples by ELISA and cross-reactivity experiments,results showed that the antigen has specific reactogenicity,which provide material source for the establishment of HDV diagnostic kit. |
| Keywords: | Hepatitis delta virus Hepatitis delta antigen prokaryotic expression ELISA |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |