Breitman M L, Tsuda M, Usukura J, Kikuchi T, Zucconi A, Khoo W, Shinohara T
Division of Molecular and Developmental Biology, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
J Biol Chem. 1991 Aug 15;266(23):15505-10.
S-antigen (S-Ag) is an abundant protein of the retina and pineal gland that elicits experimental autoimmune uveitis and pinealocytis in several animal species. To study the elements regulating the expression of S-Ag, we generated transgenic mice expressing the chloramphenicol acetyl transferase (CAT) gene under the control of a 1.3-kilobase pair 5'-flanking segment of the mouse S-Ag gene. While all of the transgenic mice expressed CAT activity in the retina, in some animals CAT activity was also detected in the pineal gland, lens, and brain. Immunoblotting, polymerase chain reaction-mediated detection of RNA, and immunocyto-staining of transgenic tissues with antibodies to CAT and S-Ag established that the profile of expression of the transgene corresponded to that of S-Ag; both proteins were detectable in retinal photoreceptor cells, pinealocytes, lens fiber and epithelial cells, the cerebellum, and the cerebral cortex. These results indicate that S-Ag is expressed in a wider spectrum of the cell types than previously recognized and that a 1.3-kilobase pair S-Ag promoter segment contains sufficient information to direct appropriate tissue-specific gene expression in transgenic mice.
S抗原(S-Ag)是视网膜和松果体中的一种丰富蛋白质,在几种动物物种中可引发实验性自身免疫性葡萄膜炎和松果体炎。为了研究调节S-Ag表达的元件,我们构建了转基因小鼠,其在小鼠S-Ag基因1.3千碱基对的5'侧翼片段控制下表达氯霉素乙酰转移酶(CAT)基因。虽然所有转基因小鼠在视网膜中均表达CAT活性,但在一些动物的松果体、晶状体和脑中也检测到了CAT活性。免疫印迹、聚合酶链反应介导的RNA检测以及用抗CAT和S-Ag抗体对转基因组织进行免疫细胞染色表明,转基因的表达谱与S-Ag的表达谱一致;两种蛋白质在视网膜光感受器细胞、松果体细胞、晶状体纤维和上皮细胞、小脑和大脑皮层中均可检测到。这些结果表明,S-Ag在比以前认识到的更广泛的细胞类型中表达,并且1.3千碱基对的S-Ag启动子片段包含足以在转基因小鼠中指导适当的组织特异性基因表达的信息。