小鼠干扰素γ（MuIFN-γ）cDNA的分子克隆及其在异源哺乳动物细胞中的表达。

Molecular cloning of murine interferon gamma (MuIFN-gamma) cDNA and its expression in heterologous mammalian cells.

作者信息

Dijkmans R, Volckaert G, Van Damme J, De Ley M, Billiau A, De Somer P

出版信息

J Interferon Res. 1985 Summer;5(3):511-20. doi: 10.1089/jir.1985.5.511.

DOI:10.1089/jir.1985.5.511

Abstract

A complete cDNA clone of murine interferon-gamma (MuIFN-gamma) was obtained by recombining two appropriate segments from partial cDNA clones originally identified by colony hybridization with rat IFN-gamma chromosomal gene fragments as probes. An expression vector was constructed in which the cDNA was placed under control of the SV40 early promoter. Transient expression of MuIFN-gamma was obtained by transformation of COS-1 cells. Subsequently, this interferon expression unit was linked to a vector containing a dihydrofolate reductase (DHFR) modular gene and used to transform DHFR(-)-CHO cells. Cell clones were selected that constitutively produce an interferon activity which by several criteria was found to be indistinguishable from natural, splenocyte-derived MuIFN-gamma.

摘要

通过将最初用大鼠干扰素 -γ 染色体基因片段作为探针经菌落杂交鉴定的部分 cDNA 克隆的两个合适片段进行重组，获得了小鼠干扰素 -γ（MuIFN -γ）的完整 cDNA 克隆。构建了一个表达载体，其中 cDNA 置于 SV40 早期启动子的控制之下。通过 COS -1 细胞的转化获得了 MuIFN -γ 的瞬时表达。随后，将这个干扰素表达单元与一个含有二氢叶酸还原酶（DHFR）模块基因的载体相连，并用于转化 DHFR(-)-CHO 细胞。选择了组成型产生干扰素活性的细胞克隆，经多项标准鉴定发现其与天然的、脾细胞来源的 MuIFN -γ 没有区别。