MOLELCULAR BIOLOGY OF PHOTOTRANSDUCTION

我们已经鉴定了人和小鼠的S抗原基因，33 K 来自小鼠和人类的蛋白质基因，以及来自人类的Shuzin基因。 的 测定人和小鼠S抗原的基因序列。 的 S抗原基因全长约50 kbp，包含16个外显子 内含子15个，占97%，外显子3%。 5'-flanking 基因的区域，大约1.5 kbp长，没有已知的调控序列。 转录元件，如TATA、GC或CCAAT盒。 5 '- 人、牛和小鼠基因表达组织的侧翼区 在体外和体内转录中的特异性启动子活性 以及在转基因小鼠中。 从人和小鼠中分离了两个33 K蛋白基因， 测定了它们的DNA序列。 他们的基因大约是10 kbp长度。含有四个外显子和三个内含子。 33 K基因 蛋白质似乎代表了一个基因家族，至少有三个 基因序列相似。 视网膜蛋白Shuzin的功能作用是未知的。 几 分离cDNA。 对人和牛的基因进行了测序， 测定了人Shuzin的全基因序列。 这种基因， 由两个内含子和三个外显子组成，具有高度重复性， 5 '-非编码区中的序列。 我们构建了含有5 '侧翼S抗原基因的融合基因， 细菌基因氯霉素乙酰基上游序列 转移酶（CAT），并显微注射含有5 '- 将小鼠S-抗原基因和CAT基因的侧翼区插入 转基因小鼠这些小鼠在视网膜中表达CAT活性， 松果体，表明1,300 bp长的S抗原启动子 片段包含足够的信息来引导适当的组织- 转基因小鼠中特异性基因表达。此外，S抗原是 发现于透镜纤维和上皮细胞、小脑和 大脑皮层 这些结果表明，S-抗原表达于 比以前认识到的更广泛的细胞类型。

We have characterized the S-antigen genes from human and mouse, 33K protein genes from mouse and human, and a Shuzin gene from human. The gene sequences of the human and mouse S-antigens were determined. The S-antigen genes were approximately 50 kbp in length, contained 16 exons and 15 introns, and comprised 97% intron and 3% exon. The 5'-flanking regions of the genes, approximately 1.5 kbp long, had no known regulatory elements for transcription such as TATA, GC, or CCAAT boxes. The 5'- flanking regions of the human, cow, and mouse genes expressed tissue specific promoter activity in both the in vitro and in vivo transcription assays as well as in transgenic mice. Two genes of the 33K protein from human and mouse were isolated, and their DNA sequences were determined. Their genes were approximately 10 kbp in length. contained four exons and three introns. The gene of 33K protein seems to represent a family of genes and to have at least three genes with similar sequences. The functional role of the retinal protein Shuzin is unknown. Several cDNA's were isolated. Those from human and cow were sequenced, and the entire gene sequence of human Shuzin was determined. This gene, which is composed of two introns and three exons, has a highly repetitive sequence in the 5'-noncoding region. We have constructed fusion genes containing a 5'-flanking S-antigen gene sequence upstream of the bacterial gene chloramphenicol acetyl transferase (CAT) and microinjected a hybrid gene containing the 5'- flanking region of the mouse S-antigen gene and the CAT gene into transgenic mice. Those mice expressed CAT activity in the retina and pineal gland, suggesting that the 1,300 bp-long S-antigen promoter segment contains sufficient information to direct appropriate tissue- specific gene expression in transgenic mice. In addition, S-antigen was found in the lens fiber and epithelial cells, the cerebellum, and the cerebral cortex. These results indicated that S-antigen is expressed in a wider spectrum of cell types than previously recognized.