Development of a bicistronic vector driven by the human polypeptide chain elongation factor 1alpha promoter for creation of stable mammalian cell lines that express very high levels of recombinant proteins.
Development of a bicistronic vector driven by the human polypeptide chain elongation factor 1alpha promoter for creation of stable mammalian cell lines that express very high levels of recombinant proteins.
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开发由人多肽链延伸因子 1α 启动子驱动的双顺反子载体，用于创建表达非常高水平的重组蛋白的稳定哺乳动物细胞系。
DOI：
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发表时间：
1998
期刊：
影响因子：
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通讯作者：
J. Wallace
中科院分区：
文献类型：
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作者：
Stephen Hobbs;S. Jitrapakdee;J. Wallace
我们开发了一种新的载体，用于在哺乳动物细胞中稳定表达重组蛋白。在这个名为 pEFIRES-P 的载体中，重组 cDNA 和嘌呤霉素抗性基因 (pac) 均被转录为由强人类多肽链延伸因子 1α 启动子驱动的单一信息。内部核糖体进入位点的存在确保任何对嘌呤霉素具有抗性的克隆也表达高水平的由 pac 基因上游克隆的 cDNA 编码的重组蛋白。我们报告了使用该载体生成表达人丙酮酸羧化酶的稳定细胞系，并表明在转染和克隆选择后，所有二十个赋予嘌呤霉素高剂量抗性的菌落也表达非常高水平的功能性丙酮酸羧化酶。使用 pEFIRES-P 可以快速有效地生成用于重组蛋白表征的稳定细胞系。
We have developed a new vector for the stable expression of recombinant protein in mammalian cells. In this vector, designated pEFIRES-P, both the recombinant cDNA and the puromycin resistant gene (pac) are transcribed as a single message driven by the strong human polypeptide chain elongation factor 1alpha promoter. The presence of an internal ribosome entry site ensures any clones that are resistant to puromycin also express high levels of recombinant protein encoded by cDNA cloned upstream from the pac gene. We report the use of this vector to generate stable cell lines expressing human pyruvate carboxylase and show that following transfection and clonal selection, all twenty colonies that conferred high dose resistance to puromycin also expressed very high levels of functional pyruvate carboxylase. Use of pEFIRES-P should permit the rapid and efficient generation of stable cell lines for the characterisation of recombinant proteins.
使用 MDR1 作为选择标记和治疗基因的双顺反子和双基因逆转录病毒载体。
影响因子：
3.7
作者：
Metz,MZ;Matsumoto,L;Winters,KA;Doroshow,JH;Kane,SE
通讯作者：
Kane,SE