摘要
目的研究TK基因在真核细胞中的表达。方法应用已构建和表达的单纯疱疹病毒I型胸苷激酶(HSV 1TK)融合蛋白免疫兔制备抗HSV 1TK血清 ,并以免疫印迹法鉴定其特异性 ,以免疫荧光法检测HSV 1TK基因在转染人肝癌细胞系中的表达。结果免疫印迹显示 ,自制和国外提供的抗TK兔血清 ,均能特异性地识别Mr 约为97000的TK融合蛋白。免疫荧光染色得到的结果满意 ,而且自制抗血清的效价和染色强度均优于国外提供的免疫血清。结论原核表达的HSV 1TK融合蛋白具有较好的免疫学活性 ;自制抗HSV
Aim To study herpes simplex virus type-1 thymidine kinase(HSV-1 TK)gene expression in eukaryotic cells. Methods The strategy included three steps:(1)Rabbit was immunized with the HSV 1 TK fusion protein(Mr 97 000);(2)The self made rabbit anti HSV 1 TK serum was analysed by Western blot and compared with the rabbit anti HSV 1 TK serum provided abroad;(3) The human hepatocellular carcinoma cell line containing HSV 1 TK gene (HHCC/TK) was examined by immunofluorescence staining with these rabbit anti HSV 1 TK sera. Results Western blot analysis showed that both the selfmade and abroad prouided rabbit anti HSV 1 TK sera could recognize a same fusion protein band. In addition, using immunofluorescence with these anti sera, HHCC/TK cells presented a clear staining. The titer and staining intensity of the self made rabbit anti HSV 1 TK serum appeared to be superior to rabbit anti HSV 1 TK serum provided abroad. Conclusion The HSV 1 TK fusion protein expressed in E.coli possesses good immunogenicity. The specific rabbit anti HSV 1 TK serum can be a useful reagent for studying HSV 1 TK gene expression in transfected cells and for cancer therapy.
出处
《细胞与分子免疫学杂志》
CSCD
2000年第4期305-307,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金部分资助!No.39570807
关键词
单纯疱疹病毒
胸苷激酶
基因治疗
基因表达
herpes simplex virus
thymidine kinase
gene
therapy
gene expression
polyclonal antibody
