[由SV40早期启动子控制的人成纤维细胞干扰素基因在CHO细胞中的组成型表达]
[Constitutive expression of human fibroblast interferon gene controlled by SV40 early promoter in CHO cells].
作者信息
Wang X M, Yu M, Wang J X, Wu S H, Hu Y W, Hou Y D
机构信息
Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing.
出版信息
Yi Chuan Xue Bao. 1989;16(4):312-7.
HindII fragment encoding human fibroblast interferon (IFN beta) gene coding sequence was fused at 60 bp downstream from the RNA start site of SV40 early gene to be a constitutive expression plasmid pSVE beta. This recombinant plasmid was transfected into the dihydrofolate reductase (dhfr)-deficient Chinese hamster ovary (CHO) cells together with a selectable dhfr gene. About half of transformants continuously secreted IFN beta into the supernatant without inducement. One of the subclone transformants constitutively produced up to 852U IFN beta/2 X 10(6) cells/ml. 48hr in common medium.
编码人成纤维细胞干扰素（IFNβ）基因编码序列的HindII片段在SV40早期基因RNA起始位点下游60 bp处融合，构建成组成型表达质粒pSVEβ。该重组质粒与一个可选择的二氢叶酸还原酶（dhfr）基因一起转染到缺乏二氢叶酸还原酶（dhfr）的中国仓鼠卵巢（CHO）细胞中。大约一半的转化体在无诱导的情况下持续将IFNβ分泌到上清液中。其中一个亚克隆转化体在普通培养基中，每2×10⁶个细胞/毫升可组成型产生高达852U的IFNβ，持续48小时。
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