摘要
从单纯疱疹病毒２型（ＨＳＶ２）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ＨＳＶ２ｇＤ基因与ＰＷＲ４５０１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。ＳＤＳＰＡＧＥ分析表明，表达产物占大肠杆菌菌体总蛋白含量的５０１％。经ＤｏｔＥＬＩＳＡ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。
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.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
1997年第3期157-159,共3页
Chinese Journal of Dermatology
基金
广东省科委自然科学基金
