论文部分内容阅读
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 1TK兔免疫血清可为进一步研究TK基因在真核细胞中的表达及用于治疗提供了一种有用的试剂
Objective To study the expression of TK gene in eukaryotic cells. Methods HSV 1TK fusion protein was used to immunize rabbits with HSV 1TK fusion protein and its specificity was identified by immunoblotting. The HSV 1TK gene was detected by immunofluorescence staining Human hepatoma cell line expression. Results Immunoblotting showed that both TK and rabbit anti-TK provided by ourselves and foreign countries could specifically recognize the TK fusion protein of Mr 97000. Immunofluorescence staining results are satisfactory, and homemade antiserum titers and staining intensity are better than the immune serum provided abroad. CONCLUSION: The prokaryotic HSV 1TK fusion protein has good immunological activity. The self-made anti-HSV 1TK rabbit serum can provide a useful reagent for the further study of TK gene expression in eukaryotic cells