Hsu T Y, Pai C Y, Shieh S M, Cho S M, Liu M Y, Chen J Y, Yang C S
Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei.
J Med Virol. 1992 Nov;38(3):214-9. doi: 10.1002/jmv.1890380311.
Two cDNA clones covering the N- and C-terminal portions of the EBV BXLF1 open reading frame were selected from a cDNA library derived from P3HR1 cells. The two clones were ligated, the N-terminal untranslated region truncated, and the product inserted into an E. coli expression vector, pET3CP*. The fusion protein was expressed under control of the T7 phage phi 10 gene promoter and shown to possess thymidine kinase activity. The protein was then used as an antigen to detect antibody reactivities in serum samples of nasopharyngeal carcinoma patients and healthy blood donors. Using a 1:400 dilution of serum samples in Western blot analyses, it was possible to differentiate the reactivities of serum IgA of NPC patients and healthy donors. The prevalence of positive reactivity to EBV TK in NPC was around 84%. The test was compared to others used for early diagnosis of NPC and was able to detect some patients who were negative in those tests.
从源自P3HR1细胞的cDNA文库中筛选出两个覆盖EBV BXLF1开放阅读框N端和C端部分的cDNA克隆。将这两个克隆连接起来，截短N端非翻译区，然后将产物插入大肠杆菌表达载体pET3CP*中。融合蛋白在T7噬菌体φ10基因启动子的控制下表达，并显示具有胸苷激酶活性。然后将该蛋白用作抗原，以检测鼻咽癌患者和健康献血者血清样本中的抗体反应性。在蛋白质印迹分析中使用1:400稀释的血清样本，可以区分鼻咽癌患者和健康供体血清IgA的反应性。鼻咽癌患者中对EBV TK呈阳性反应的比例约为84%。将该检测方法与其他用于鼻咽癌早期诊断的方法进行比较，能够检测出在那些检测中呈阴性的一些患者。
