Herpes simplex virus expressing Epstein-Barr virus nuclear antigen 1.
Herpes simplex virus expressing Epstein-Barr virus nuclear antigen 1.
复制标题
表达 Epstein-Barr 病毒核抗原 1 的单纯疱疹病毒。
DOI：
10.1016/0042-6822(86)90330-2
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发表时间：
1986
期刊：
影响因子：
3.7
通讯作者：
Kieff,E
中科院分区：
文献类型：
--
作者：
Hummel,M;Arsenakis,M;Marchini,A;Lee,L;Roizman,B;Kieff,E
将含有已知编码EB病毒（EBV）的大部分或全部EBNA 1蛋白的开放阅读框的DNA片段以适当的转录方向融合到启动子调节结构域，加帽位点。和单纯疱疹病毒1型（HSV-1）HSV-1 α4基因5′端转录非编码序列的一部分。在这些构建体中，20、130或385 bp的EBV DNA和28 bp的HSV-1 DNA将α4帽位点与EBNA 1基因的假定起始密码子分开。将嵌合α4-EBNA 1基因导入L细胞或利用胸苷激酶选择系统重组到病毒基因组中。存在于L细胞克隆中的三种嵌合基因构建体表达的蛋白质在电泳和免疫学性质方面与真实EBNA 1无区别，表明EBV开放阅读框开始处的ATG启动了真正EBNA 1蛋白的翻译。存在于L细胞中的嵌合α 4 EBNA 1基因被HSV-1感染诱导，并被调节为α基因。重组到病毒基因组中的嵌合α4-EBNA 1基因也作为α基因被调控。重组病毒是稳定的，并且表达的EBNAl是携带EBV基因组的人淋巴细胞中通常表达的EBNAl的50至100倍。EBNA 1不改变HSV-1蛋白表达程序。讨论了该矢量的实用性。
DNA fragments containing an open reading frame known to encode most or all of the EBNAl protein of Epstein-Barr virus (EBV) were fused in the proper transcriptional orientation to the promoter regulatory domain, capping site. and a portion of the 5′ transcribed noncoding sequences of the HSV-1 α4 gene of herpes simplex virus 1 (HSV-1). In these constructs 20, 130, or 385 bp of EBV DNA and 28 bp of HSV-1 DNA separated the α4 cap site from a putative initiator codon of the EBNA1 gene. The chimeric α4-EBNA1 genes were introduced into L cells or recombined into the viral genome using the thymidine kinase selection system. The three chimeric gene constructs resident in the L cell clones expressed a protein indistinguishable from authentic EBNA1 with respect to electrophoretic and immunologic properties indicating that the ATG at the beginning of the EBV open reading frame initiated translation of the bonafide EBNAl protein. The chimeric α4EBNA1 genes resident in L cells were induced by HSV-1 infection and were regulated as α genes. The chimeric α4-EBNA1 gene recombined into the viral genome was also regulated as an α gene. The recombinant viruses were stable and expressed 50- to 100-fold more EBNAl than is ordinarily expressed in human lymphocytes carrying the EBV genome. EBNA1 did not alter the program of HSV-1 protein expression. The utility of the vector is discussed.