ENHANCER-DEPENDENT EXPRESSION OF HUMAN KAPPA-IMMUNOGLOBULIN GENES INTRODUCED INTO MOUSE PRE-B LYMPHOCYTES BY ELECTROPORATION
ENHANCER-DEPENDENT EXPRESSION OF HUMAN KAPPA-IMMUNOGLOBULIN GENES INTRODUCED INTO MOUSE PRE-B LYMPHOCYTES BY ELECTROPORATION
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DOI：
10.1073/pnas.81.22.7161
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
LEDER, P
中科院分区：
文献类型：
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作者：
POTTER, H;WEIR, L;LEDER, P
一种将克隆基因引入哺乳动物细胞的通用方法被开发出来，这种方法比目前的技术提供了实质性的好处。它简单，快速，适用于许多（也许是所有）细胞类型，包括那些难以传统转染程序的细胞。该方法包括将细胞悬浮液和克隆DNA暴露在高压放电中。在该转染程序的模型应用中，克隆的人和小鼠Ig .kappa的表达。基因稳定导入小鼠前b细胞和成纤维细胞。在人kappa的J-C内含子中存在一个b细胞特异性增强激活子区。在小鼠前b淋巴细胞中有效转录克隆基因所必需的基因。这表明DNA元件及其调控活性所需的蛋白质在进化过程中高度保守，并且这些元件在免疫细胞发育的前b细胞阶段起作用，这一阶段先于产生。基因重排。
A general method for introducing cloned genes into mammalian cells that affords substantial benefits over current technology was developed. It is simple, rapid and applicable to many (perhaps all) cell types, including those that are refractory to traditional transfection procedures. The method involves exposure of a suspension of cells and cloned DNA to a high-voltage electric discharge. In a model application of this transfection procedure, the expression of cloned human and mouse Ig .kappa. genes stably introduced into mouse pre-B cells and fibroblasts. There is a B-cell-specific enhancer-activator region in the J-C intron of the human .kappa. gene that is necessary for efficient transcription of the cloned gene in mouse pre-B lymphocytes. This suggests that both the DNA element and the proteins required for its regulatory activity have been highly conserved in evolution and that these elements operate at the pre-B-cell stage of immunocyte development, a stage that precedes productive .kappa. gene rearrangement.