Walls D, Perricaudet M, Gannon F
Department of Microbiology, University College, Galway, Ireland.
Nucleic Acids Res. 1988 Apr 11;16(7):2859-72. doi: 10.1093/nar/16.7.2859.
We have used small random EBV B95-8 DNA fragments to generate a large genomic bank in a plasmid expression vector. This bank was screened with a pool of sera from individuals with IM thus allowing any EBV antigen which evoked an immune response in man to be identified. The characterization of four immunopositive clones obtained in this way is presented in this study. Three of these clones express viral ORF DNA sequences which are parts of larger ORFs in the BamH1 N(het), V and X regions of the B95-8 viral genome. cDNA cloning has been used to confirm that the cloned sequences from BamH1 N and V are expressed in cell culture and to identify the transcription units involved. The fourth clone expresses an ORF sequence located in the viral BamH1 F fragment in a region not previously recognized as having protein coding potential. The experimental design used here must reflect the situation in vivo and consequently these sequences must be expressed and be antigenic during IM.
我们使用小的随机EBV B95-8 DNA片段在质粒表达载体中构建了一个大型基因组文库。用传染性单核细胞增多症患者的血清池筛选该文库，从而能够鉴定出任何在人体内引发免疫反应的EBV抗原。本研究展示了以此方式获得的四个免疫阳性克隆的特征。其中三个克隆表达病毒ORF DNA序列，这些序列是B95-8病毒基因组BamH1 N（het）、V和X区域中较大ORF的一部分。已使用cDNA克隆来确认来自BamH1 N和V的克隆序列在细胞培养中表达，并鉴定所涉及的转录单位。第四个克隆表达位于病毒BamH1 F片段中的一个ORF序列，该区域以前未被认为具有蛋白质编码潜力。这里使用的实验设计必须反映体内情况，因此这些序列在传染性单核细胞增多症期间必须表达且具有抗原性。
