利用大肠杆菌recA启动子表达人干扰素基因。
Expression of human interferon genes using the recA promoter of Escherichia coli.
作者信息
Feinstein S I, Chernajovsky Y, Chen L, Maroteaux L, Mory Y
出版信息
Nucleic Acids Res. 1983 May 11;11(9):2927-41. doi: 10.1093/nar/11.9.2927.
Interferon beta 1 and three alpha-interferon genes were cloned on Eco RI fragments isolated from a human genomic library into the Eco RI site of a plasmid containing the recA promoter of E. coli. Expression of interferon activity from cells carrying these plasmids was nalidixic acid inducible. The alpha-interferon genes were expressed only when in the same transcriptional orientation as the recA promoter while the beta 1 interferon gene was expressed in either orientation. Interferon activity was also inducibly expressed from the recA promoter in cells containing a plasmid carrying a fusion of the recA gene with the beta 1 interferon gene. This interferon activity was thirty-fold less sensitive to neutralization by polyclonal antibodies than authentic interferon, implying that the change near the amino terminus affects either antibody recognition or specific activity or both.
β-干扰素1和三种α-干扰素基因被克隆到从人基因组文库分离的Eco RI片段上，该片段插入到含有大肠杆菌recA启动子的质粒的Eco RI位点。携带这些质粒的细胞中干扰素活性的表达可被萘啶酸诱导。α-干扰素基因只有在与recA启动子处于相同转录方向时才表达，而β1干扰素基因在任何方向均可表达。在含有携带recA基因与β1干扰素基因融合体的质粒的细胞中，recA启动子也可诱导表达干扰素活性。这种干扰素活性对多克隆抗体中和作用的敏感性比天然干扰素低30倍，这意味着氨基末端附近的变化影响抗体识别或比活性或两者。
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