发明授权
US5908763A DNA encoding GM-CSF and a method of producing GM-CSF protein
失效
编码GM-CSF的DNA和生产GM-CSF蛋白的方法
- 专利标题: DNA encoding GM-CSF and a method of producing GM-CSF protein
- 专利标题(中): 编码GM-CSF的DNA和生产GM-CSF蛋白的方法
-
申请号: US287019申请日: 1994-08-08
-
公开(公告)号: US5908763A公开(公告)日: 1999-06-01
- 发明人: Steven C. Clark , Randal J. Kaufman , Gordon G. Wong , Elizabeth A. Wang
- 申请人: Steven C. Clark , Randal J. Kaufman , Gordon G. Wong , Elizabeth A. Wang
- 申请人地址: CHX Basel
- 专利权人: Novartis Corporation
- 当前专利权人: Novartis Corporation
- 当前专利权人地址: CHX Basel
- 主分类号: A61K38/00
- IPC分类号: A61K38/00 ; C07K14/535 ; C12N15/27 ; C12N15/70 ; C12N15/81 ; C12N15/85 ; C07K14/53 ; C12N15/63
摘要:
A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.
公开/授权文献
信息查询
IPC分类: