EBNA3 GENES OF EBV

EBV 的 EBNA3 基因

基本信息

批准号：
6132316
负责人：
KARA L CARTER
金额：
$ 3.84万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-10-01 至
项目状态：
未结题

项目摘要

The function of the Epstein-Barr virus nuclear antigen (EBNA) 3 genes will be analyzed. This approach will be a three-fold effort. First, genetic analysis of these genes will be preformed by generating a series of deletional and insertional mutants and assessing the effect of these mutations on the ability of the recombinant viruses to transform primary B lymphocytes as wild-type EBV does. Second, a search for cellular proteins that interact with the EBNA 3 proteins will be undertaken by using the yeast two-hybrid system and fusion-protein pull-down experiments. Once these proteins are identified, the function significance of their interactions with the EBNA 3 proteins will be analyzed by altering the ability of these interactions to occur in the cell and determining the effect on transformation of primary B lymphocytes. Finally, because the EBNA 3 proteins are nuclear proteins and have been shown to interact with RBJk, a cellular DNA binding protein, the induction of cellular genes by these proteins will be studied. To do this, cell lines that express each of the EBNA 3 protein independently will be compared to EBV infected cell lines and noninfected cell lines by differential display analysis. The cDNA's for the cellular genes induced by the EBNA 3 proteins will be cloned and analyzed.

EB 病毒核抗原 (EBNA) 3 基因的功能将进行分析。这种方法将是三倍的努力。第一、遗传对这些基因的分析将通过生成一系列缺失和插入突变体并评估这些突变体的影响突变对重组病毒转化原代细胞能力的影响 B 淋巴细胞与野生型 EBV 一样。二、搜索蜂窝与 EBNA 3 蛋白相互作用的蛋白将由使用酵母双杂交系统和融合蛋白下拉实验。一旦这些蛋白质被识别出来，其功能它们与 EBNA 3 蛋白相互作用的重要性将是通过改变这些相互作用发生的能力来分析细胞并确定对原代 B 转化的影响淋巴细胞。最后，因为 EBNA 3 蛋白是核蛋白并已被证明与细胞 DNA 结合蛋白 RBJk 相互作用，将研究这些蛋白质对细胞基因的诱导。要做的事这是独立表达每种 EBNA 3 蛋白的细胞系将与 EBV 感染细胞系和未感染细胞系进行比较差异显示分析。诱导的细胞基因的 cDNA 通过 EBNA 3 蛋白质将被克隆和分析。