论文部分内容阅读
３０℃培养基因工程菌，迅速转移至４２℃培养４ｈ诱导重组质粒中外源ＥＢ病毒胸苷激酶（ＥＢＶＴＫ）基因的表达。菌体蛋白作十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）分析表明，重组蛋白分子大小与ＥＢＶＴＫ分子大小的理论值６７ｋｕ相符，占菌体总蛋白的６％以上。Ｗｅｓｔｅｒｎｂｌｏｔ分析表明重组蛋白可与混合ＮＰＣ病人血清中ＥＢＶＴＫ特异性抗体反应，呈现良好的抗原性。以３ＨｄＴ为底物测定菌体总蛋白的ＴＫ比活性，结果提示，重组蛋白具有胸苷激酶活性。研究为ＥＢＶＴＫ在ＮＰＣ诊断中的应用打下了基础。
30 ℃ cultured engineered bacteria, rapidly transferred to 42 ℃ for 4h to induce expression of exogenous EBV thymidine kinase (EBVTK) gene in the recombinant plasmids. SDS-PAGE analysis showed that the size of the recombinant protein corresponded to 67ku of the theoretical size of the EBVTK molecule, accounting for more than 6% of the total bacterial protein. Western blot analysis showed that the recombinant protein reacted with EBVTK-specific antibody in the serum of patients with mixed NPC and showed good antigenicity. The TK specific activity of total bacterial proteins was determined by 3HdT. The results showed that the recombinant protein has thymidine kinase activity. The research laid the foundation for the application of EBVTK in the diagnosis of NPC.