BCL8, A NOVEL GENE REARRANGED IN DLLC

这个项目的目标是在1511-13年描述一种新的基因 命名为BCL8，它与DLLC中的IgH基因重排。这个 提出了以下具体目标。1.基因组和c DNA的组织 并预测了BCL8的蛋白。该基因以多种形式表达 组织，特别是在非淋巴组织中，如前列腺和睾丸。 利用睾丸cDNA文库，我们克隆了BCL8的cDNA.我们现在就会 BCL8基因组和cDNAs的组织结构测定及开发 针对适合检测基因的预测蛋白的抗血清 Western blotting和免疫组织化学方法检测其表达。2.机制 在DLLC中放松对BCL8的管制。我们将确定以下后果： DLLC中bcl8基因失控引起的染色体易位。由北方 杂交分析，我们将确定bcl8的频率和水平 B细胞DLLC中的表达及其与BCL8重排的关系 我们还将调查是否有更多放松监管的机制，例如 5‘调控区的突变或缺失起作用。通过 用BCL8探针和3‘、5’和3‘进行常规基因组克隆 RACE(快速扩增cDNA末端)技术，我们将识别 BCL8与染色体易位连接的序列 14q32以外的站点。这样的分析将澄清机制(S)和 影响bcl8的错配易位的后果。3.政府的角色 Bcl8在正常和淋巴组织肿瘤的发生发展中的作用。我们将调查 Bcl8基因异常表达在淋巴样肿瘤发生发展中的作用 将含BCL8基因的表达载体导入EB病毒载体 永生化的B淋巴母细胞系。表达载体包含 在免疫球蛋白基因增强子调控下的bcl8基因将用于 构建转基因小鼠在B细胞中高效表达转基因 血统。这些研究将阐明bcl8在正常人中的作用。 发育和淋巴样肿瘤。

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.