2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/37996Epstein-Barr virus (EBV) is a gamma herpesvirus, which is persistent in B lymphoid cells, enhances transformation and tumorigenesis of latently infected B cells in vitro via LMP-induced cellular survival protein, Bcl-2. The effect of Bcl-2 protein on B cell survival obviously plays a key role. Recently, an early lytic gene, BALF1, which is homologous to Bcl-2 protein in structure, is characterized in EBV infected cells. However, the role and function of BALF1 in EBV infected cells is still to be in confusion and be controversial. In this proposal, we first intend to introduce a BALF1 expression vector to cells and to see the effect of BALF1 on the promoters of latent genes, LMP and EBNA1 or immediate early genes, Z promoter. In the meantime, transfection of a pBALF1-siRNA plasmid, which suppresses the expression of BALF1, into cells is to see whether withdrawal of BALF1 from cells will change the cell survival state or cause cell apoptotsis. In addition, we will employ cell human cellular cDNA microarray to see which cellular genes affected by BALF1. The results of microarray assay may speculate the effectiveness of BALF1 on infected cells, according to the BALF1 regulated cellular genes. Furthermore, we plan to generate a BALF1 mutant recombinant EBV to test the effect of BALF1 on EBV other genes, to evaluate the effect on cell transformation and on cell cycle genes. Finally we will demonstrate the effect of BALF1 recombinant EBV on cell apoptosis. From this study, we probably acquire information of BALF1 that what function BALF1 is in infected cells and how to regulate the relationship between EBV and host cells. Understanding the function of BALF1 in infected cells will facilitate the development of drugs to inhibit its intrinsic activities and non-immunogenic vectors for use in human gene therapy.EB病毒是屬於皰療病毒科(herpesviridae)—員，它能夠持續存活於B淋巴細胞，並透 過其潛伏性膜蛋白1 (LMP1)活化細胞中的Bcl-2蛋白質。最後致使B細胞轉形 (transformation)及癌化°從此現象顯示Bcl-2蛋白在其中扮演重要角色°近來研冗發現 EB病毒中，有一蛋白質會出現在溶裂期的早期稱爲BALF1。其蛋白結構與Bcl-2相似， 但對於它在細胞中的功能則有爭議。爲了釐清此點，在本計劃中首先建構BALF1蛋白 表現質体，送入細胞中觀察其對細胞及病毒基因的影響。同時建構另一個具有抑制 BALF1表現的質体(pBALFl-siRNA)。並觀察其送入細胞後對細胞所產生的影響。此外 我們利用人類的cDNA微距陣(microarray)，去推測BALF1如何調控細胞中的基因， 以達成改變細胞的狀態。進一步我們試圖製造一株BALF1基因突變的EB病毒。利用 此突變病毒株自然感染進入細胞後，觀察其與野生株(wildtype)對細胞基因及其存活是 否有影響。另外並觀察其是否調控其他病毒基因表現。從本次研究計劃，我們可以從 中獲得BALF1基因對其寄主及病毒本身調控機制的資訊。而從資訊中又可找出宿主與 病毒互動的關係。並透過此層深入的了解，進而發展出抑制EB病毒的藥物及基因治療 病毒所產生的疾病方法。Project