中华皮肤科杂志 ›› 1997, Vol. 30 ›› Issue (3): 157-159.

• 论著 • 上一篇    下一篇

疱疹病毒gD2基因在大肠杆菌中的高效表达

杨慧兰1, 李明2, 廖元兴1, 江悦华2, 吴锦银2, 徐广坤3   

  1. 1. 广州军区总医院皮肤科, 510010;
    2. 广州军区总医院医学实验科, 510010;
    3. 中山医科大学附属第三医院皮肤科
  • 收稿日期:1996-08-06 修回日期:1996-12-24 出版日期:1997-06-15 发布日期:1997-06-15
  • 基金资助:
    广东省科委自然科学基金

High Level Expression of Glycoprotein D of Herpes Simplex Virus in Escherichia Coli

Yang Huilan1, Li Ming2, Liao Yuanxing1   

  1. Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010
  • Received:1996-08-06 Revised:1996-12-24 Online:1997-06-15 Published:1997-06-15

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摘要： 从单纯疱疹病毒2型(HSV-2)Sav株中扩增糖蛋白D(gD)基因646bp片段,克隆入M13mp18、M13mp19测序载体。将经测序证明的HSV-2gD基因与PWR450-1表达载体进行重组,在大肠杆菌中表达了β-半乳糖苷酶-gD融合蛋白。SDS-PAGE分析表明,表达产物占大肠杆菌菌体总蛋白含量的50.1%.经Dot-ELISA及免疫印迹鉴定证实表达产物具有免疫学活性。对gD2表达产物进行了初步纯化,纯度达90%.gD2的高效表达和纯化有助于研究其生物学功能和免疫学特性,为研制生殖器疱疹基因工程疫苗和免疫学诊断试剂提供有利条件。

关键词: 单纯疱疹病毒, 糖蛋白D

Abstract: Herpes simplex virus glycoprotien D gene was amplified from HSV-2 Sav strain. The D gene fragment was cloned into M13mp18 and M13mp19 and sequenced.The gD and expression vector(PWR450-1) were recombined, the recombinants could express high level corresponding beta galactosidase antigen fusion proteins in E.coli.The SDS-PAGE analysis demonstrated that the expressed fusion protein consists of 50.1% of total bacterial proteins. The immunogenicity of the product was tested with dot-ELISA and Western blot. Preliminary purification of gD galactosidase fusion protein showed that the product exhibied a purity over 90%.The results of investigation will help to study the immunogenicity of gD and produce genetic engineering vaccine of genital herpes simplex.

Key words: Herpes simplex virus, Glycoprotein D