摘要
利用RT-PCR技术从人A431细胞中克隆表皮生长因子受体(epidermalgrowthfactorreceptor,EGFR)基因N端序列,经测序鉴定正确。将膜外序列不同功能及模拟抗原呈递较佳的亚克隆DNA片段插入T7噬菌体表达载体,从而构建了重组T7噬菌体抗肿瘤疫苗并用于动物免疫注射,用斑点印迹(dotblot)和MTT法分别检测到特异性抗体和细胞毒性T淋巴细胞反应。结果显示,以噬菌体颗粒为载体的疫苗可以诱导体液免疫反应和细胞免疫反应。
The overexpression of epidermical growth factor receptor （EGFR） played a vital role in the process of the formation, progression, metastasis and therapy of the malignant tumor. The monoclonal antibodys have been used for treatment of EGFR-positive expression tumors in clinical trial. In our study the N-terminal of EGFR gene was cloned from human A431 cells with RT-PCR technology and the sequence was identified. The subclone DNA fragments with different function and better presentation of mimic antigen were inserted into the bacteriophage T7 expression vector. The anti-cancer vaccine of recombinant bacteriophages T7 were constructed and the mice were immunized. The specific antibodies and the cytotoxic T lymphocyte response were detected with dot blot and MTT method, respectively. The result showed that recombinant bacteriophage T7 carder vaccine could induced humoral immune reaction and cellular immune reaction. Therefore, the recombinant bacteriophage T7 carrier could be a kind of promising vaccine and provides a basis for the study of anti-cancer vaccine.
出处
《细胞生物学杂志》
CSCD
2006年第2期217-222,共6页
Chinese Journal of Cell Biology
基金
上海市科委立项资助重点课题(No.0319236)~~
