Ohno T, Wang X, Kurashima J, Saijo-Kurita K, Hirono M
RIKEN Cell Bank, RIKEN Institute of Physical and Chemical Research, Tsukuba Science City, Japan.
Cytotechnology. 1991 Nov;7(3):165-72. doi: 10.1007/BF00365927.
We have established a novel cell line from a Vero cell derivative that is useful for expression of exogenous genes and protein production. Parental Vero-317 cells can grow in biotin-containing Eagle's MEM without supplements. By transforming this cell line with replication origin-defective SV40 DNA, which contains a temperature-sensitive tsA58 large T antigen gene, we established the Verots S3 cell line that amplified a SV40-origin containing plasmid. The cell line expressed a human growth hormone (hGH) gene insert with higher efficiency than COS-7 cells in 5% serum-containing MEM and could grow and continue hGH expression in protein-free MEM. However, temperature-sensitive shut down of hGH production was observed not immediately but 3 days after the temperature shift from 33 degrees C to 39.5 degrees C.
我们从一种Vero细胞衍生物中建立了一种新型细胞系，该细胞系可用于外源基因的表达和蛋白质生产。亲代Vero-317细胞可以在含有生物素的Eagle's MEM中生长，无需添加其他成分。通过用含有温度敏感型tsA58大T抗原基因的复制起点缺陷型SV40 DNA转化该细胞系，我们建立了Verots S3细胞系，该细胞系可扩增含有SV40起点的质粒。在含5%血清的MEM中，该细胞系表达人生长激素（hGH）基因插入片段的效率高于COS-7细胞，并且可以在无蛋白的MEM中生长并持续表达hGH。然而，hGH产量的温度敏感型关闭不是在温度从33℃转变为39.5℃后立即观察到，而是在3天后观察到。
