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弹性蛋白酶-人生长激素融合基因在转基因小鼠胰腺腺泡细胞中的特异性表达。

Specific expression of an elastase-human growth hormone fusion gene in pancreatic acinar cells of transgenic mice.

作者信息

Ornitz D M, Palmiter R D, Hammer R E, Brinster R L, Swift G H, MacDonald R J

出版信息

Nature. 1985;313(6003):600-2. doi: 10.1038/313600a0.

Abstract

Transfection of genes into tissue culture cell lines has demonstrated that relatively short DNA sequences can allow expression of immunoglobulin, insulin and chymotrypsin genes in their appropriate cell types. A definitive test of cell-specific gene expression, however, requires testing genes in every possible cell type, an experiment performed easily by introducing the gene in question into the germ line of an animal. Transfer of intact genes into mice has demonstrated that a mouse immunoglobulin kappa gene is expressed specifically in B lymphocytes, a rat elastase I gene is expressed specifically in pancreas and a chicken transferrin gene is expressed preferentially in liver. Mouse metallothionein-growth hormone fusion genes introduced into mice are preferentially expressed in the liver, consistent with the expression of endogenous metallothionein genes, but initial experiments with beta-globin genes have not revealed proper regulation. To identify the DNA elements required for pancreas-specific expression of the rat elastase I gene, we joined the 5'-flanking region of this gene to the human growth hormone (hGH) structural gene and introduced the fusion gene into mice. Here we demonstrate that a fusion gene containing only 213 base pairs (bp) of elastase I gene sequence directs expression of hGH in pancreatic acinar cells.

摘要

将基因转染到组织培养细胞系中已表明,相对较短的DNA序列能够使免疫球蛋白、胰岛素和胰凝乳蛋白酶基因在其相应的细胞类型中表达。然而,对细胞特异性基因表达的确定性测试需要在每一种可能的细胞类型中对基因进行检测,而通过将相关基因导入动物的生殖系中就能轻松完成这样的实验。将完整基因导入小鼠体内已表明,小鼠免疫球蛋白κ基因在B淋巴细胞中特异性表达,大鼠弹性蛋白酶I基因在胰腺中特异性表达,鸡运铁蛋白基因在肝脏中优先表达。导入小鼠体内的小鼠金属硫蛋白-生长激素融合基因在肝脏中优先表达,这与内源性金属硫蛋白基因的表达情况一致,但β-珠蛋白基因的初步实验尚未揭示出正常的调控机制。为了确定大鼠弹性蛋白酶I基因胰腺特异性表达所需的DNA元件,我们将该基因的5'侧翼区与人生长激素(hGH)结构基因连接起来,并将融合基因导入小鼠体内。在此我们证明,一个仅包含213个碱基对(bp)弹性蛋白酶I基因序列的融合基因能够指导hGH在胰腺腺泡细胞中表达。

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