用负向选择增富法筛选人GM-CSF cDNA  

Isolation, identification and expression of a cDNA clone encoding human GM-CSF

作　　者：孙英[1] 马大龙[1] 劳者歌 庞建 宋泉声[1] 尉达民 税朝祥[2] 陈萍生 王良绪[2] 杨景山 唐岩[3] 

机构地区：[1]北京医科大学免疫学教研室,北京100083 [2]北京医科大学第三医院血液研究室,北京100083 [3]北京医科大学组胚教研室,北京100083

出　　处：《中国免疫学杂志》1990年第4期247-249,共3页Chinese Journal of Immunology

摘　　要：用负向选择增富法成功地获得人GM-CSF cDNA克隆,筛选阳性率提高至1/170。用该cDNA转染的CDS-7细胞上清可测出集落刺激活性。经亚克隆化后在大肠杆菌表达出重组人GMCSF,可维持细胞株BMU-2的生长并促进急性非淋巴细胞白血病(ANLL)患者骨髓细胞的集落形成。One of the human GM-CSF cDNA was successfully isolated with a new screening method. Up 1/170 of positive rate was reached. The COS-7 cells were transfected with the cDNA clone and the resulting supernants contained colony-stimulatory activities. Human GM-CSF cDNA was also subcloned into a expression plasmid pKPL-2D and the unfused recombinant human GM-CSF was expressed in E. coli. The expressed products could maintain in vitro the growth of a cell line BMU- 2 and stimulate its colony formation. In addition, it could increase the colony formation of bone marrow cells from the patients with acute non-lymphatic leukemia (ANLL).

关 键 词：GM-CSF CDNA 地高辛 探针 表达 

分 类 号：R392－33[医药卫生—免疫学]