Biggin M, Farrell P J, Barrell B G
EMBO J. 1984 May;3(5):1083-90. doi: 10.1002/j.1460-2075.1984.tb01933.x.
The sequence of the BamHI L fragment of B95-8 Epstein-Barr virus (EBV) DNA has been determined. The transcription starts of five promoters have been mapped to this fragment, using S1 mapping and either in vitro transcription or the primer extension technique. Dramatically increased levels of cytoplasmic poly(A) + RNAs, transcribed from these promoters, occur after treatment of B95-8 cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). For three of these promoters addition of phosphonoacetic acid (PAA) inhibits the effect of TPA, indicating that they give rise to late lytic cycle RNAs. The other two promoters give rise to early RNAs. Northern blot analysis indicates that one of the late promoters initiates two transcripts whose size differences are due to different splicing patterns. These two RNAs code for the 350/300 and 250/200 kd envelope glycoproteins of EBV. The sequences of these proteins would be of use in the production of a synthetic vaccine to prevent EBV infection.
已确定了B95 - 8爱泼斯坦 - 巴尔病毒(EBV)DNA的BamHI L片段序列。利用S1核酸酶图谱分析以及体外转录或引物延伸技术,已将五个启动子的转录起始位点定位到该片段上。在用12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)处理B95 - 8细胞后,从这些启动子转录的细胞质多聚腺苷酸(poly(A)+)RNA水平显著增加。对于其中三个启动子,添加膦甲酸(PAA)可抑制TPA的作用,这表明它们产生晚期裂解周期RNA。另外两个启动子产生早期RNA。Northern印迹分析表明,其中一个晚期启动子起始两种转录本,其大小差异归因于不同的剪接模式。这两种RNA编码EBV的350/300和250/200 kd包膜糖蛋白。这些蛋白的序列将有助于生产预防EBV感染的合成疫苗。