ALCOHOL METABOLISM GENES IN TRANSGENIC MICE

描述：肝脏中的乙醇代谢由I类介导 乙醇脱氢酶（ADH）和细胞色素P4502 E1（CYP 2 E1）。I类ADH 在小鼠中由Adh-1基因编码，该基因在复杂的激素和 发展控制转基因小鼠将被用来确定作用 基因内和5 '-和3'-侧翼序列控制的总 表达表型与ADH相连的延伸至约10 kb的区域 minigene控制肾脏、肾上腺和其他组织中的适当表达; 然而，肝脏表达需要其它顺式作用元件。 含有所有基因内序列的粘粒和λ基因组克隆， 不同的5 '-和3'-侧翼距离现在被隔离并且将被使用 在转基因分析中， 表情相同的转基因方法，使用替代启动子， 粘粒克隆，将用于产生过表达ADH的转基因小鼠 和/或CYP 2 E1。一种独特的分析方法将允许RNA和蛋白质 从转基因产生的，以区别于从转基因产生的， 内源基因对于Adh-1，所使用的独特遗传变体将 能够在组织内的细胞类型水平上正确表达， 测定过表达小鼠将被用于测试主要假设 乙醇引起的肝损伤包括自由基形成、脂质过氧化 过氧化和脂肪肝的产生。此外，一种遗传变异 在近交系中鉴定的Adh-3表达将在 分子水平，并将提供一种机制， 这个基因的表达。

DESCRIPTION: Ethanol metabolism in the liver is mediated by class I alcohol dehydrogenase (ADH) and cytochrome P4502E1 (CYP2E1). Class I ADH in mice is encoded by the Adh-1 gene which is under complex hormonal and developmental control. Transgenic mice will be used to determine the role of intragenic and 5'- and 3'-flanking sequences in control of the total expression phenotype. A region extending to -10kb attached tot an ADH minigene controls proper expression in kidney, adrenal, and other tissues; however, other cis-acting elements are required for liver expression. Cosmid and lambda genomic clones containing all intragenic sequences and different 5'- and 3'-flanking distance are now isolated and will be used in transgenic analysis to identify sequences necessary for liver expression. The same transgenic approach, using alternative promoters and cosmid clones, will be used to produce transgenic mice over expressing ADH and/or CYP2E1. A unique approach to analysis will allow RNA and protein produced from the transgene to be distinguished from production from the endogenous gene. For Adh-1, the unique genetic variant to be used will enable proper expression at the level of cell type within a tissue to be determined. Over expressing mice will be use to test major hypotheses about ethanol-induced liver injury including free radical formation, lipid peroxidation and production of fatty-liver. In addition, a genetic variant in Adh-3 expression identified among inbred strains will be defined at the molecular level and will provide a mechanism for differential tissue expression of this gene.