EBV INTERNAL REPEAT TRANSCRIPTION UNIT IN LATENCY

EB病毒与伯基特淋巴瘤有关， 未分化鼻咽癌，以及一些 淋巴组织增生性疾病 在体外病毒可以将B 淋巴细胞转化为含有附加体的永生淋巴母细胞系 病毒基因组并沿病毒基因的沿着子集表达。 基因组由 独特的短（12 kb）和长（124 kb）区域，由30 kb的大区域分隔 3.1kb序列的串联重复序列测定（IR 1）。 IR 1是主要的物理 基因组的特征，在潜伏期期间转录，编码其中一种 潜伏核抗原（EBNA 4，也称为EBNA-LP），并有助于 所有EBNA的mRNA的前导外显子。 复杂的拼接模式 在潜伏期内， 并且在从潜伏状态诱导病毒产生之后， 通过直接北方印迹与 跨越剪接点的寡核苷酸和聚合酶链反应 使用与以下互补的特异性引物扩增mRNA片段： 剪接点和外显子，以及用于3 ′和5 ′锚的非特异性引物。 由于PCR只需要少量的初始材料， 将发展直接检查组织活检技术。 的 在体外实验中，通过添加处理， 印刷受体. 所获得的信息将广泛映射IR 1 转录单位，开发用于检查IR 1控制的体外模型 转录加工，并定义IR 1对原位转录的贡献。 潜伏期和淋巴瘤发生。 开发的技术和试剂将 可用于进一步的研究，也可用于诊断 评估EBV相关淋巴增生性疾病的发病机制 和肿瘤。

The Epstein-Barr virus is associated with Burkitt's lymphoma, undifferentiated nasopharyngeal carcinoma, and a number of lymphoproliferative disorders. In vitro the virus can convert B lymphocytes into immortal lymphoblastoid cell lines which contain episomal viral genomes and express along subset of viral genes. The genome consists of unique short (12kb) and long (124kb) regions separated by a large 30kb tandem repeat assay (IR1) of a 3.1kb sequence. IR1 is the major physical feature of the genome, is transcribed during latency, encodes one of the latent nuclear antigens (EBNA4, also termed the EBNA-LP) and contributes leader exons to the mRNAs of all the EBNAs. The complex splicing pattern of latent transcripts through IR1 upon initial infection, during latency and after induction of virus production from a latent state will be investigated by a combination of direct Northern blotting with olignucleotides spanning splice junctions and polymerase chain reaction amplification of mRNA fragments using specific primers complementary to splice junctions and exons, and non-specific primers for 3' and 5' anchors. Since only small amounts of initial material are necessary for PCR, this technique will be developed for direct examination of tissue biopsies. The mechanism of regulation will be dissected in vitro by adding processing substrates. The information obtained will extensively map the IR1 transcription unit, develop in vitro models for examining control of IR1 transcript processing, and define the contribution of IR1 to in situ latency and lymphomagenesis. The techniques and reagents developed will be useful in further studies and may also be of diagnostic utility in assessing the pathogenesis of EBV associated lymphoproliferative disorders and neoplasms.