Gross T G, Sakai K, Volsky D J
Biochem Biophys Res Commun. 1986 Feb 13;134(3):1260-8. doi: 10.1016/0006-291x(86)90386-4.
Fresh human lymphocytes were transfected with cloned EBV DNA fragments containing the coding exon for EBNA-1. DNA-loaded reconstituted Sendai Virus envelopes (RSVE) were used for efficient gene transfer. EBNA was detected by immunofluorescence in 1-5% of cells transfected with either the cloned BamHl K fragment of EBV DNA (5.1 kb) or the recombinant plasmid pSV3neoEBNA1, containing only the 2.0kb EBNA-1 coding exon. EBV-specific mRNA was detected by hybridization up to 14 days after DNA transfer. Quantitation of mRNA by laser densitometry revealed that the transcription level was similar to that obtained after infection with an intact virus. However, no effect on cellular proliferation was observed by (3H)-thymidine incorporation assays, and transformation was not achieved. We conclude that though EBNA-1 may be necessary for cellular immortalization by EBV, it alone is not sufficient.
用含有EBNA-1编码外显子的克隆EBV DNA片段转染新鲜的人淋巴细胞。加载了DNA的重组仙台病毒包膜（RSVE）用于高效基因转移。在用EBV DNA的克隆BamHl K片段（5.1 kb）或仅含有2.0 kb EBNA-1编码外显子的重组质粒pSV3neoEBNA1转染的1%-5%的细胞中，通过免疫荧光检测到EBNA。在DNA转移后长达14天通过杂交检测到EBV特异性mRNA。通过激光密度测定法定量mRNA显示转录水平与用完整病毒感染后获得的水平相似。然而，通过（3H）-胸苷掺入试验未观察到对细胞增殖的影响，并且未实现转化。我们得出结论，虽然EBNA-1可能是EBV使细胞永生化所必需的，但仅它本身是不够的。
