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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

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【作者】 孙英; 马大龙; 劳者歌; 庞建; 宋泉声; 尉达民; 税朝祥; 陈萍生; 王良绪; 杨景山; 唐岩;

【Author】 Sun Ying, et al. Department of Immunology, Beijing Medical University, Beijing 100083, China

【机构】 北京医科大学免疫学教研室; 北京医科大学第三医院血液研究室; 北京医科大学组胚教研室; 北京医科大学组胚教研室 北京 100083; 北京 100083; 北京 100083;

【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。

【Abstract】 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; 地高辛素探针; 表达;
【Key words】 GM-CSF; cDNA; Expression;
  • 【文献出处】 中国免疫学杂志 ,Chinese Journal of Immunology , 编辑部邮箱 ,1990年04期
  • 【被引频次】1
  • 【下载频次】15
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