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目的：构建高效表达EBVTK的重组克隆体系。方法：以pUC8X为模板，5’-CTGAATTCATGGCTGGATTTCC-3’及5’CAACTGCAGCCTAGTCCCGATT-3’为引物，用PCR技术扩增出EBV tk基因的DNA片段。EcoR I／Pst I 双酶切PCR产物和载体pBV220，T4连接酶使目的基因tk定向克隆至选定质粒pBV220中。结果：重组质粒pBV220-tk经EcoR I／Pst I双酶切后得两条电泳带分别位于1．8 kb、3．6 kb处；以pBV220-tk为模板，两引物同上进行PCR扩增，产物电泳带于1．8 kb处。结论：目的基因tk已定向克隆至质粒pBV220中“,”Objective：To construct a recombinant plasmid which can express large amounts of the Epstein-Barr-virus-coded thymidine kinase．Methods：The plasmid PUC8X was used as a template for PCR with the 5’primer GTGAATTCATGGCTGGATTTCC and the 3’primer CAACTGCAGCCTAGTCCCGATT to generate a fragment with a 5’EcoR I site and a 3’Pst I site．The DNA was cut to produce an EcoR I-Pst I fragment that was ligated into the similarly cut vector pBV220 with T4 ligase．Results：The recombmant DNA pBV220-tk was constructed，which was cut by EcoR I and Pst I and analyzed by agarose gel electrophoresis．Two bands at 1．8 kb and 3．6 kb were obtained．The pBV220-tk was used as a template for PCR with two primers indicated above，the PCR-generated fragment was at 1．8 kb analyzed by agarose gel electrophoresis．Conclusion：The aimed gene tk has been inserted into the pBV220 successfully．