REGULATION OF GENE EXPRESSION IN THE ADENOVIRUS SYSTEM AND IN TRANSGENIC MICE

我们的实验室研究哺乳动物细胞中的基因控制机制 在文化和整个动物。 两大项目之一， 具有E1 A调控功能的腺病毒，已经完成。 的 E1 A功能作为一种转录调节因子， 恶性转化 编码E1 A蛋白或某些 这些蛋白质的结构域已被插入原核表达中， 载体，并在E.杆菌 我们显微注射了 这些蛋白质进入哺乳动物细胞，并测量它们迁移到 细胞核并激活腺病毒E1 A缺失突变体。 核定位和病毒基因激活的信息， 已显示由腺病毒E1 A基因的不同结构域编码。 我们的第二个项目涉及将特定的基因构建体插入到 小鼠胚胎 表达的空间和时间控制 在得到的转基因动物中检查插入的基因。 到目前为止， 我们分析了携带三种不同嵌合基因构建体的小鼠。 每一个都含有细菌氯霉素乙酰基的基因 转移酶（CAT）的启动子/增强子区域的控制下， 来自小鼠α 2（I）胶原基因或来自劳斯肉瘤病毒 （RSV）。 携带病毒小鼠体内CAT表达的时空调控 α A堆叠-CAT或α 2（I）胶原CAT构建体反映了 真正的小鼠基因的5'侧翼序列来自该基因 得到嵌合基因。 在携带RSV-CAT构建体的小鼠中， CAT表达优先针对肌肉和结缔组织 组织. 这特别反映了肉瘤病毒的疾病。 最后，我们已经开始分析一个RSV-CAT转基因株， 以胚胎致死的显性特征为特点。

Our laboratory investigates mechanisms of gene control in mammalian cells in culture and in the whole animal. One of two major projects, dealing with the E1A regulatory function of adenovirus, has been completed. The E1A function acts as a transcriptional modulator and is involved in malignant transformation. Sequences encoding E1A proteins or certain domains of these proteins have been inserted in prokaryotic expression vectors, and E1A proteins have been produced in E. coli. We microinjected the proteins into mammalian cells and measured their ability to migrate to the cell nucleus and to activate an adenovirus E1A deletion mutant. Information for nuclear localization and for viral gene activation has been shown to be encoded by distinct domains of the adenovirus E1A gene. Our second project involves insertion of specific gene constructs into mouse embryos. Spatial and temporal control of the expression of the inserted genes is examined in the resulting transgenic animals. So far, we have analyzed mice carrying three difference chimeric gene constructs. Each of these contains the gene for bacterial chloramphenicol acetyl transferase (CAT) under the control of a promoter/enhancer region derived from either the mouse Alpha2(I) collagen gene or from Rous sarcoma virus (RSV). The temporal and spatial contol of CAT expression in mice carrying the AlphaA crystallin-CAT or the Alpha2(I) collagen CAT construct reflects that of the genuine mouse genes from which the 5' flanking sequences of the chimeric genes were derived. In mice carrying the RSV-CAT construct CAT expression is preferentially directed to muscle and connective tissue. This reflects the disease specifically of sarcoma viruses. Finally, we have begun to analyze one RSV-CAT transgenic strain which is characterized by a dominant trait of embryonic lethality.