Littler E, Zeuthen J, McBride A A, Trøst Sørensen E, Powell K L, Walsh-Arrand J E, Arrand J R
EMBO J. 1986 Aug;5(8):1959-66. doi: 10.1002/j.1460-2075.1986.tb04450.x.
We have demonstrated the presence of an Epstein-Barr virus (EBV)-coded thymidine kinase (TK) by producing biochemically transformed, TK-positive mammalian cell lines using either microinjection of whole EBV virions or calcium phosphate-mediated transfection of the SalI-B restriction endonuclease fragment of EBV DNA. Analysis of these cell lines showed that: (i) EBV DNA was present in the cell lines, (ii) sequences from the SalI-B restriction endonuclease fragment of EBV were expressed, (iii) a TK activity was present and (iv) a protein with antigenic cross-reactivity with the herpes simplex virus (HSV) TK was produced. The identity of the EBV TK gene was determined by demonstrating that a recombinant plasmid, which expressed the protein product of the BXLF1 open reading frame as a fusion protein, could complement TK- strains of E. coli. A comparison of the predicted amino acid sequences of the TK proteins of EBV and HSV-1 revealed significant regions of homology.
我们通过使用全EBV病毒粒子显微注射或EBV DNA的SalI - B限制性内切酶片段的磷酸钙介导转染来产生生化转化的、TK阳性的哺乳动物细胞系，从而证明了爱泼斯坦 - 巴尔病毒（EBV）编码的胸苷激酶（TK）的存在。对这些细胞系的分析表明：（i）细胞系中存在EBV DNA，（ii）EBV的SalI - B限制性内切酶片段的序列得以表达，（iii）存在TK活性，以及（iv）产生了一种与单纯疱疹病毒（HSV）TK具有抗原交叉反应性的蛋白质。通过证明一个表达BXLF1开放阅读框的蛋白质产物作为融合蛋白的重组质粒能够互补大肠杆菌的TK - 菌株，确定了EBV TK基因的身份。EBV和HSV - 1的TK蛋白预测氨基酸序列的比较揭示了显著的同源区域。
