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CLONING AND DEVELOPMENT OF MONKEY TRANSFORMATION GROWTH FACTOR BETA 1

机译：猴转化生长因子β1的克隆与发展

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摘要

Recombinant transforming growth factor- beta 1 (TGF- beta 1) is expressed to high levels in Chinese hamster ovary (CHO) cells using dihydrofolate reductase (dhfr) gene amplification. The expression plasmid was derived from the pSV2 vectors and contained, in tandem, the simian TGF- beta 1 and mouse dhfr cDNAs. Transcription of both cDNAs was controlled by the SV40 early promoter. Stepwise selection of transfected CHO cells in increasing concentrations of methotrexate yielded cell lines expressing amplified TGF- beta 1 nucleic acid sequences. The expression plasmid DNA was amplified greater than 35-fold in one of the methotrexate selected transfectants. The major proteins secreted by these cells consisted of latent TGF- beta 1 and TGF- beta 1 precursor polypeptides as judged by immunoblots using site-specific anti-peptide antibodies derived from various regions of the TGF- beta 1 precursor. Levels of recombinant TGF- beta 1 protein secreted by these cells approached 30 ug/24 hour/107 cells.

机译：使用二氢叶酸还原酶（dhfr）基因扩增，在中国仓鼠卵巢（CHO）细胞中高水平表达重组转化生长因子-β1（TGF-β1）。表达质粒衍生自pSV2载体，并串联包含猿猴TGF-β1和小鼠dhfr cDNA。 SV40早期启动子控制两个cDNA的转录。以递增浓度的甲氨蝶呤逐步选择转染的CHO细胞，可产生表达扩增的TGF-β1核酸序列的细胞系。在选择的甲氨蝶呤转染子之一中将表达质粒DNA扩增大于35倍。这些细胞分泌的主要蛋白质由潜在的TGF-β1和TGF-β1前体多肽组成，通过免疫印迹使用衍生自TGF-β1前体各个区域的位点特异性抗肽抗体进行判断。这些细胞分泌的重组TGF-β1蛋白的水平接近30ug / 24小时/ 10 7细胞。

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公开/公告号JPH01257497A

专利类型
公开/公告日1989-10-13

原文格式PDF
申请/专利权人 ONCOGEN LP;
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申请/专利号JP19880130501
发明设计人 ANSONII EFU PARUCHIO;RARII JIENTORII;DANIERU TOWAADOJITSUKU;
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申请日1988-05-30
分类号C12N15/09;A61K38/00;A61P35/00;C07K1/20;C07K14/00;C07K14/495;C07K14/52;C12N5/10;C12N15/85;C12P21/02;C12R1/91;
国家 JP
入库时间 2022-08-22 06:47:19

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