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 在文化和整个动物中。两个主要项目之一，处理 具有E1A腺病毒的调控功能，已经完成。这个 E1a功能作为转录调节器参与了 恶变。编码E1A蛋白或某些特定蛋白的序列 这些蛋白的结构域已插入原核表达 载体，在大肠杆菌中表达了E1a蛋白。我们进行了显微注射 这些蛋白质进入哺乳动物细胞，并测量它们迁移到 并激活腺病毒E1a缺失突变体。 核定位和病毒基因激活的信息有 已被证明是由腺病毒E1A基因的不同结构域编码的。 我们的第二个项目涉及将特定的基因结构插入到 小鼠胚胎。基因表达的时空调控 在产生的转基因动物中检查插入的基因。到目前为止, 我们已经分析了携带三种不同嵌合基因结构的小鼠。 其中每一种都含有细菌氯霉素乙酰基的基因。 启动子/增强子区控制下的转移酶(CAT) 来自小鼠α2(I)胶原基因或来自Rous肉瘤病毒 (RSV)。荷瘤小鼠CAT基因表达的时空调控 AlphaA晶体蛋白-CAT或Alpha2(I)胶原CAT构建反映 这是真正的小鼠基因，来自5‘侧翼序列 获得了嵌合基因。在携带RSV-CAT结构的小鼠中 CAT的表达主要针对肌肉和结缔组织 组织。这反映了这种疾病对肉瘤病毒的特异性。 最后，我们已经开始分析一株RSV-CAT转基因株 以胚胎致死的显性特征为特征。

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.