Mueller-Lantzsch N, Lenoir G M, Sauter M, Takaki K, Béchet J M, Kuklik-Roos C, Wunderlich D, Bornkamm G W
EMBO J. 1985 Jul;4(7):1805-11. doi: 10.1002/j.1460-2075.1985.tb03854.x.
Cell lines were established by co-transfection of cloned M-ABA Epstein-Barr virus (EBV) DNA fragments with plasmids conferring resistance to dominant selective markers. A baby hamster kidney cell line carrying the HindIII-I1 fragment exhibits a nuclear antigen of 82 000 daltons, serologically defined as EBV-determined nuclear antigen (EBNA) 1. Furthermore, a Rat-1 cell line transfected with DNA of the clone pM 780-28 containing three large internal repeats (BglII-U) and the adjacent BglII-C fragment expresses a nuclear antigen of 82 000 daltons which can be visualized only by a subset of anti EBNA-positive human sera. Sera recognizing the 82 000-dalton protein of the transfected cell line reacted with a protein of the same size in the non-producer line Raji, designated as EBNA 2. Conversely, sera without reactivity to the 82 000-dalton protein failed to react with EBNA 2 of Raji cells. P3HR-1 and Daudi cells with large deletions in BglII-U and -C are devoid of EBNA 2. The data presented provide evidence that a second EBNA protein is encoded by the region of the EBV genome which is deleted in the non-transforming P3HR-1 strain.
通过将克隆的M-ABA Epstein-Barr病毒（EBV）DNA片段与赋予对显性选择标记抗性的质粒共转染来建立细胞系。携带HindIII-I1片段的幼仓鼠肾细胞系表现出一种82000道尔顿的核抗原，血清学上定义为EBV确定的核抗原（EBNA）1。此外，用含有三个大的内部重复序列（BglII-U）和相邻的BglII-C片段的克隆pM 780-28的DNA转染的大鼠1细胞系表达一种82000道尔顿的核抗原，只有一部分抗EBNA阳性的人血清才能检测到它。识别转染细胞系中82000道尔顿蛋白质的血清与非产生细胞系Raji中相同大小的蛋白质发生反应，该蛋白质被指定为EBNA 2。相反，如果血清对82000道尔顿的蛋白质没有反应，那么它也不会与Raji细胞的EBNA 2发生反应。在BglII-U和-C区域有大片段缺失的P3HR-1和Daudi细胞缺乏EBNA 2。本文提供的数据表明，EBV基因组中在非转化性P3HR-1菌株中缺失的区域编码了第二种EBNA蛋白。
