Epstein-Barr病毒胸苷激酶在大肠杆菌表达的研究

EXPRESSION OF THYMIDINE KINASE FOR EPSTEIN-BARR VIRUS IN E.COLI

研究重组 EB病毒胸苷激酶 (thym idine kinase of Epstein- Barr virus,EBV TK)在大肠杆菌中的表达。将 EB病毒的 tk基因与原核表达载体 p BV2 2 0进行重组 ,构成质粒 p BV2 2 0 / tk,并在大肠杆菌 DH5α中获得表达 ;采用限制性内切酶分析进行重组质粒的鉴定、 SDS- PAEG电泳和 Western blot反应检测蛋白的表达。结果显示成功地构建了 p BV2 2 0 / tk质粒 ,并有重组蛋白的表达 ,该重组蛋白可与鼻咽癌病人血清发生特异性反应。结果表明该重组蛋白可作为抗原应用于鼻咽癌病人血清 TK Ig A抗体的检测 ,为重组 TK的生产。

To study the expression of thymidine kinase for Epstein Barr virus in E.coli, the recombinant prokaryotic expression plasmid pBV220/tk was constructed by inserting the thymidine kinase fragment of Epstein Barr virus (EBV) into pBV220 and expressed in E.coli DH5α. The recombinant plasmid pBV220/tk was identified with restriction enzyme analysis, and the recombinant protein with SDS PAEG electrophoresis and Western blot assay. It was found that the recombinant pBV220/tk was constructed successfully, and the recombinant protein expressed in E.coli. Furthermore the protein showed specific reaction to the serum of patients with nasopharyngeal carcinoma (NPC). These results indicate that the recombinant protein can be used as an antigen for the detection of thymidine kinase IgA antibody in the serum of patients with NPC, and our research has provided experimental evidence for its production and clinical use.