Mizukami T, Itoh S
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd.
J Biochem. 1987 May;101(5):1307-10. doi: 10.1093/oxfordjournals.jbchem.a121995.
A useful vector, pAGE103, has been developed for the expression of cDNA in animal cells using the simian virus 40 (SV40) expression signals. cDNA could be expressed easily by inserting it into the multiple cloning sites (HindIII, SalI/AccI, XbaI, BamHI, SmaI/XmaI, KpnI/Asp718, SacI and EcoRI) of the vector, which are located between the SV40 early promoter and the SV40 early RNA processing signals for splicing and polyadenylation. In addition to the above transcription unit, pAGE103 contains the replication origin of ColE1, and a dual KmR/G418R selective gene. Several unique restriction sites are located on the boundaries between the above-mentioned three components of the vector, allowing the easy substitution or insertion of other genetic elements. The human interferon-beta gene was inserted into pAGE103 and shown to be expressed transiently in COS-1 cells and stably in several animal cell lines.
已开发出一种有用的载体pAGE103,用于利用猿猴病毒40(SV40)表达信号在动物细胞中表达cDNA。通过将cDNA插入载体的多克隆位点(HindIII、SalI/AccI、XbaI、BamHI、SmaI/XmaI、KpnI/Asp718、SacI和EcoRI),可轻松实现cDNA的表达,这些多克隆位点位于SV40早期启动子与用于剪接和聚腺苷酸化的SV40早期RNA加工信号之间。除上述转录单元外,pAGE103还包含ColE1的复制起点以及一个双重KmR/G418R选择基因。几个独特的限制性酶切位点位于载体上述三个组分之间的边界上,便于其他遗传元件的轻松替换或插入。人β干扰素基因被插入到pAGE103中,并显示在COS-1细胞中瞬时表达,在几种动物细胞系中稳定表达。