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通过DNA介导的基因转移实现Thy-1糖蛋白基因的表达。

Expression of the Thy-1 glycoprotein gene by DNA-mediated gene transfer.

作者信息

Evans G A, Ingraham H A, Lewis K, Cunningham K, Seki T, Moriuchi T, Chang H C, Silver J, Hyman R

出版信息

Proc Natl Acad Sci U S A. 1984 Sep;81(17):5532-6. doi: 10.1073/pnas.81.17.5532.

Abstract

We isolated a gene encoding the Thy-1.2 glycoprotein from a recombinant library constructed from BALB/c mouse DNA. To evaluate the expression of this cloned gene in different genomic environments, we introduced it into cell lines derived from fibroblast, lymphoid, and neuronal tissues by DNA-mediated gene transfer. When integrated into the genome of mouse L cells, cell-surface Thy-1 can be detected with anti-Thy-1 monoclonal antibodies. These L-cell lines contain between two and four copies of the cloned Thy-1 gene stably integrated in the host genome. After subcloning into a plasmid vector containing the bacterial Eco-gpt gene as a selectable marker, the Thy-1 gene was introduced into the Thy-1-deficient mouse lymphoma AKR1 (Thy-1-d), and the rat neuronal cell line, B50. The resulting transformants also contain two to four copies of the cloned Thy-1 gene but express up to 50-fold more cell-surface Thy-1.2 than the L cell transformants. The expression of vastly differing amounts of cell-surface Thy-1 from similar numbers of genes suggests that the gene encoding this differentiation antigen is under tissue-specific regulation.

摘要

我们从由BALB/c小鼠DNA构建的重组文库中分离出一个编码Thy-1.2糖蛋白的基因。为了评估该克隆基因在不同基因组环境中的表达情况,我们通过DNA介导的基因转移将其导入源自成纤维细胞、淋巴组织和神经组织的细胞系中。当整合到小鼠L细胞的基因组中时,可用抗Thy-1单克隆抗体检测到细胞表面的Thy-1。这些L细胞系含有稳定整合在宿主基因组中的两到四个克隆的Thy-1基因拷贝。在亚克隆到含有细菌Eco-gpt基因作为选择标记的质粒载体中后,将Thy-1基因导入Thy-1缺陷型小鼠淋巴瘤AKR1(Thy-1-d)和大鼠神经细胞系B50中。所得的转化体也含有两到四个克隆的Thy-1基因拷贝,但细胞表面Thy-1.2的表达量比L细胞转化体高50倍。从相似数量的基因中表达出数量差异极大的细胞表面Thy-1,这表明编码这种分化抗原的基因受到组织特异性调控。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f08b/391740/d1fab4ee691c/pnas00618-0260-a.jpg

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