Post L E, Norrild B, Simpson T, Roizman B
Mol Cell Biol. 1982 Mar;2(3):233-40. doi: 10.1128/mcb.2.3.233-240.1982.
We are describing a system for the introduction, selection, and expression of eucaryotic genes in higher eucaryotic cells. The carrier consisted of the herpes simplex virus 1 (HSV-1) tk gene covalently linked to an HSV-1 alpha promoter directed away from the tk gene. In this study we fused to the alpha promoter the 5' transcribed noncoding sequences and the coding sequences of the chicken oviduct ovalbumin gene. Cells converted to the TK+ phenotype with this chimeric fragment produced an ovalbumin precursor which was processed and secreted into the extracellular fluid. The ovalbumin gene utilized the HSV-1 alpha promoter and was regulated as a viral gene inasmuch as inversion of the genomic DNA relative to the alpha promoter resulted in no ovalbumin synthesis, and production of ovalbumin was enhanced after superinfection with HSV-1. Synthesis of ovalbumin was not detected when cDNA was linked to the HSV-1 alpha promoter. The carrier system described in this study is suitable for introduction, selection, and expression of eucaryotic genes whose natural promoter is either weak or requires the presence of regulatory elements which may be absent from undifferentiated cells in culture.
我们正在描述一种用于在高等真核细胞中导入、选择和表达真核基因的系统。载体由与远离tk基因的单纯疱疹病毒1(HSV-1)α启动子共价连接的单纯疱疹病毒1(HSV-1)tk基因组成。在本研究中,我们将鸡输卵管卵清蛋白基因的5'转录非编码序列和编码序列与α启动子融合。用这个嵌合片段转化为TK+表型的细胞产生了一种卵清蛋白前体,该前体被加工并分泌到细胞外液中。卵清蛋白基因利用HSV-1α启动子,并作为病毒基因进行调控,因为相对于α启动子的基因组DNA倒转导致没有卵清蛋白合成,并且在HSV-1超感染后卵清蛋白的产生增强。当cDNA与HSV-1α启动子连接时,未检测到卵清蛋白的合成。本研究中描述的载体系统适用于导入、选择和表达天然启动子较弱或需要存在培养中未分化细胞可能不存在的调控元件的真核基因。