BCL8, A NOVEL GENE REARRANGED IN DLLC

这个项目的目标是在1511-13描述一个新的基因 称为BCL 8，其在DLLC中与IGH基因重排。 的 提出了以下具体目标。 1.基因组和cDNA组织 和预测的BCL 8蛋白。 该基因以多种形式表达 组织，特别是非淋巴组织，如前列腺和睾丸。 利用睾丸cDNA文库，我们克隆了BCL 8 cDNA。 我们现在将 确定BCL 8的基因组和cDNA组织， 适用于检测基因的预测蛋白质的抗血清 Western blotting和免疫组化检测表达。 2.机制 DLLC中BCL 8失调的可能性。 我们将决定 DLLC中染色体易位导致BCL 8失调。 由北方 印迹分析，我们将确定BCL 8的频率和水平 在B细胞DLLC中的表达及其与BCL 8重排的相关性。 我们还将调查是否有其他放松管制的机制，如 5 ′调控区中的突变或缺失起作用。 通过 使用BCL 8探针和3'、5'和3' RACE（cDNA末端快速扩增）技术，我们将鉴定 来自涉及BCL 8与染色体的易位连接的序列 14 q32以外的其他网站。 这种分析将澄清机制， 影响BCL 8的混杂易位的结果。 3. 作用 BCL 8在正常和淋巴肿瘤发展中的作用 我们将调查 BCL 8异常表达在淋巴肿瘤发展中的作用。 将含有BCL 8基因的表达载体转染到EBV-1中， 永生化B淋巴母细胞样细胞系。 表达载体 在IG基因增强子调控下的BCL 8基因将用于 构建转基因小鼠以在B细胞中过表达转基因 脉 这些研究将阐明BCL-8在正常人中的作用。 发展和淋巴瘤形成。

The goal of this project is to characterize a novel gene at 1511-13 termed BCL8, which rearranges with the IGH gene in DLLC. The following Specific Aims are proposed. 1. Genomic and cDNA organization and predicted protein of BCL8. The gene is expressed in multiple tissues, notably in nonlymphoid tissues such as prostate and testis. Using a testis cDNA library, we have cloned BCL8 cDNA. We will now determine the genomic and cDNA organization of BCL8 and develop antisera against the predicted protein suitable for detecting gene expression by Western blotting and immunohistochemistry. 2. Mechanism of BCL8 deregulation in DLLC. We will determine the consequence of chromosome translocation to BCL8 deregulation in DLLC. By Northern blot analysis, we will determine the frequency and level of BCL8 expression in B-cell DLLC and its correlation with BCL8 rearrangements. We will also investigate if additional mechanisms of deregulation, such as mutations or deletions n the 5' regulatory region operate. By conventional genomic cloning using BCL8 probes and 3' and 5' and 3' RACE (rapid amplification of cDNA ends) techniques, we will identify sequences from tanslocation junctions involving BCL8 with chromosomal sites other than 14q32. Such analysis will clarify the mechanism(s) and consequence of promiscuous translocations affecting BCL8. 3. Role of BCL8 in normal and lymphoid tumor development. We will investigate the role of aberrant BCL8 expression in lymphoid tumor development. Expression vectors containing BCL8 gene will be transfected into EBV- immortalized B-lymphoblastoid cell lines. Expression vectors containing the BCL8 gene under the regulation of IG-gene enhancer will be used to construct transgenic mice to overexpress of the transgene in B-cell lineage. These studies will elucidate the role of BCL-8 in normal development and lymphoid neoplasia.