影响人γ—型干扰素基因在工程菌中表达效率的因素研究

Investigation of impact of some factors affecting the expression of human recombinant IFN-γ gene in E.coil

本工作研究了影响人γ—型干扰素基因在工程菌中表达效率的3种因素；结果表明：采用M9＋LB发酵培养基，控制工程菌培养时间和温度，即种子菌在LB（Amp）液中30℃生长10h，转入发酵培养基中30℃扩增3h，42℃诱导培养3h，并按种子菌液与发酵培养液2％的比例扩增工程菌，可使发酵菌液中工程菌密度达OD（600）1．6，IFN－γ表达水平占菌体总蛋白的65％～70％，使IFN－γ活性达1．68×107IU／ml。

In this work,impact of some factors affecting the expression of human recombinant IFN-γ gene in E. coil (JF1125)were investigated. It was found that fermentable medium(LB+M9) was suitable for cell growth and IFN-γ expression. In fermentable process it was favourable that E. coil JF1125 carrying recombinant IFN-γ gene grew for 10h at 30℃ in LB then amplified for 3h at 30℃ in the fermentable medium and incubated the culture for a further 3h at 42℃. The fermentable medium is fifty times as volume as primary culture. In these coditions the level of expression and activity of IFN-γ could be reached to 1. 6×107IU/ml,about 65%~70% of total protein.