Expression and excretion of human fibroblast beta 1 interferon in monkey cells after transfection with a recombinant SV40 plasmid vector.

D Gheysen, W Fiers
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Abstract

We have constructed a eukaryotic expression vector designed to express a gene under late SV40 transcriptional control. From this chimeric plasmic-SV40 vector, virtually all the sequences which code for the major capsid protein VP1 have been deleted and instead, the human fibroblast interferon beta 1 cDNA gene has been inserted. After transfection of monkey cells with this recombinant, substantial quantities of human beta 1 interferon (up to 2 x 10(-4) IU/ml) were excreted in the culture medium. Transfection of nonpermissive mouse L cells or rat cells yielded virtually undetectable quantities of human beta 1 interferon (5 x 10(3) to 10(4) times less than that in monkey cells). The recombinant SV40 vector may serve as a model vehicle for the efficient expression of other eukaryotic genes and might also be used as a direct screening vector for cloning of eukaryotic or prokaryotic cDNA genes.

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重组SV40质粒载体转染猴细胞后人成纤维细胞β 1干扰素的表达和分泌。
我们构建了一个真核表达载体,旨在表达SV40晚期转录控制下的基因。从这个嵌合质质- sv40载体中,几乎所有编码主要衣壳蛋白VP1的序列都被删除了,取而代之的是插入了人成纤维细胞干扰素β 1 cDNA基因。用这种重组蛋白转染猴细胞后,大量的人β 1干扰素(高达2 × 10(-4) IU/ml)在培养基中排泄。转染非受纳小鼠L细胞或大鼠细胞产生的人β 1干扰素几乎无法检测到(比猴子细胞少5 × 10(3)至10(4)倍)。重组载体SV40可作为其他真核基因高效表达的模型载体,也可作为真核或原核cDNA基因克隆的直接筛选载体。
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