Stable expression in mouse cells of nuclear neoantigen after transfer of a 3.4-megadalton cloned fragment of Epstein-Barr virus DNA.
Stable expression in mouse cells of nuclear neoantigen after transfer of a 3.4-megadalton cloned fragment of Epstein-Barr virus DNA.
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转移 3.4 兆道尔顿 Epstein-Barr 病毒 DNA 克隆片段后，小鼠细胞中核新抗原稳定表达。
DOI：
10.1073/pnas.79.18.5688
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发表时间：
1982
影响因子：
11.1
通讯作者：
Miller,G
中科院分区：
文献类型：
--
作者：
Summers,WP;Grogan,EA;Shedd,D;Robert,M;Liu,CR;Miller,G
所有携带EB病毒（EBV）基因组的细胞都在细胞核中含有新抗原（EBNA）。通过转染，我们定位了编码或诱导与EBNA血清学相关的抗原的基因组片段。在EBV DNA的3.4兆道尔顿BamHI片段K中发现了负责基因，特别是在由HindIII片段I1代表的左侧1.9兆道尔顿中。用含有单纯疱疹病毒胸苷激酶基因和EB病毒DNA的EcoRI片段B或BamHI片段的重组质粒共转化小鼠LTK-细胞。将在选择性培养基中存活的TK+细胞克隆。约50%的克隆在每个细胞核中表达新抗原。这些小鼠细胞被用作免疫荧光试验中的抗原。在已知含有EBNA抗体的30份人血清中发现了核抗原抗体;在18份没有EBNA抗体的血清中未检测到。作为获得克隆的EBV DNA片段的结果，表达EBNA的小鼠细胞应证明可用于表征该抗原的结构，并作为诊断EBV感染的试剂。
All cells that harbor the Epstein-Barr virus (EBV) genome contain a neoantigen in the nucleus (EBNA). By transfection we located a segment of the genome that encodes or induces an antigen serologically related to EBNA. The responsible genes are found in the 3.4-megaldalton BamHI fragment K of EBV DNA, specifically in the left 1.9 megadaltons represented by HindIII fragment I1. Mouse LTK- cells were cotransformed with recombinant plasmids, containing the herpes simplex virus thymidine kinase gene and either EcoRI fragment B or BamHI fragment of K of EBV DNA. The TK+ cells surviving in selective medium were cloned. About 50% of the clones expressed the neoantigen in every nucleus. These mouse cells were used as antigens in immunofluorescence tests. Antibody to the nuclear antigen was found in 30 human sera known to contain antibody to EBNA; it was not detected in 18 sera that did not have antibody to EBNA. Mouse cells expressing EBNA as the result of acquisition of cloned EBV DNA fragments should prove useful in the characterization of the structure of this antigen and as reagents for the diagnosis of EBV infections.