人粒细胞-巨噬细胞集落刺激因子在大肠杆菌中的克隆与表达

Molecular Cloning and Expression of Human Granulocyte-macrophage Colony stimulating factor in Escherichia coli System

利用基因重组技术构建人粒细胞－巨噬细胞集落刺激因子（ｈＧＭ－ＣＳＦ）的高效表达菌株Ｅ．ｃｏｌｉＨＢ１０１／ｐＺＷ．ＧＭ４７，经酶切电泳、ＤＮＡ测序、ＳＤＳ－ＰＡＧＥ、Ｗｅｓｔｅｒｎ印迹及生物活性测定等分析鉴定，证明能特异性表达有生物活性的１４ｋＤａＧＭ－ＣＳＦ，表达水平达４０％以上，比活性高达５×１０＾７ｕ／ｍｇ，具有良好的开发应用前景。

Human granulocyte-macrophage stimulating factor (hGM-CSF) is an important hematopoietic growth factor. hGM-CSF is able to stimulate hematopoietic progenitors to produce granulocytes, macrophages and eosinophils. hGM-CSF also shows the potential to treat myeloid leukemias. In order to prepare enough amount of hGM-CSF. a genetically engineered strain E. coll HB101/pZW. GM47 was constructed by DNA recombination technologes. and characterized by restriction analysis. DNA sequencing, SDS-PAGE, Western blotting and bioassays. The strain has been shown to produce a 14kDa mature hGM-CSF protein with experession level higher than 40% and a specific activity of 5.0×107u/mg . It shows a good potential of clinical applications.