粒/巨噬细胞集落刺激生长因子的基因克隆表达

Gene Cloning and Expressing of GranulocyteMacrophage ColonyStimulating Factor

利用逆转录聚合酶链反应从人外周血淋巴细胞克隆获得全长ｃＤＡＮ，将ｃＤＮＡ片段克隆至质粒表达载体ＰＢＶ２２１内，转化大肠杆菌ＤＨ５α筛选获得了高表达人粒／巨噬细胞集落刺激生长因子的工程菌。表达的人粒／巨噬细胞集落刺激生长因子经ＳｅｐｈａｃｒｙｌＳ２００分子筛纯化及ＱＳｅｐｈａｒｏｓｅ阴离子交换纯化，获得了高纯度的粒／巨噬细胞集落刺激生长因子。

The cDNA of granulocytemacrophage colonystimulating factor (GMCSF) was cloned from human peripheral blood lymphocyte by RTPCR,and cloned to the expressing plasmid PBV 221.The recombinant plasmid was transformed into E.coli.DH5 for expressing GMCSF,the expressed GMCSF was about 22% of the total E.coli.protein.After purified by Sephacryl S200 chromatography and QSepharose ion exchange,the expressed GMCSF is very pure(95%) and has excellent biological activity.