Onomichi K, Eto Y, Shibai H
Central Research Laboratories, Ajinomoto Co., Inc., Kanagawa.
J Biochem. 1987 Jul;102(1):123-31. doi: 10.1093/oxfordjournals.jbchem.a122023.
Expression of human interleukin 2 (IL-2) at high levels has been achieved in Chinese hamster ovary (CHO) cells by amplification of transfected sequences. Plasmids containing the human IL-2 cDNA or genomic DNA and mouse dihydrofolate reductase (DHFR) cDNA were transfected into DHFR-negative CHO cells. Transformants expressing DHFR were selected in media lacking nucleosides, and cells which amplified both DHFR and IL-2 genes were obtained by exposure to increasing methotrexate (MTX) concentrations. These cell lines constitutively expressed elevated levels of IL-2 at a concentration of 2 mg/liter. These cell lines continued to produce IL-2 stably through at least 1 month, even in the absence of MTX.
通过转染序列的扩增,已在中国仓鼠卵巢(CHO)细胞中实现了人白细胞介素2(IL-2)的高水平表达。将含有人类IL-2 cDNA或基因组DNA以及小鼠二氢叶酸还原酶(DHFR)cDNA的质粒转染到DHFR阴性的CHO细胞中。在缺乏核苷的培养基中选择表达DHFR的转化体,并通过暴露于不断增加的甲氨蝶呤(MTX)浓度获得同时扩增DHFR和IL-2基因的细胞。这些细胞系以2毫克/升的浓度组成型表达升高水平的IL-2。这些细胞系即使在没有MTX的情况下也能稳定产生IL-2至少1个月。