发明名称 |
PRODUCTION AND PURIFICATION OF LYMPHOKINE |
摘要 |
<p>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; and selecting 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.</p> |
申请公布号 |
JPH06319557(A) |
申请公布日期 |
1994.11.22 |
申请号 |
JP19930176039 |
申请日期 |
1993.06.22 |
申请人 |
SANDOZ AG |
发明人 |
SUTEIIBUN SHII KURAAKU;RANDARU JIEI KAUFUMAN;GOODON JII UONGU;ERIZABESU EE UANGU |
分类号 |
A61K35/74;A61K35/12;A61K38/00;A61K38/16;C07H21/04;C07K1/16;C07K1/20;C07K1/36;C07K14/00;C07K14/435;C07K14/52;C07K14/53;C07K14/535;C12N;C12N1/19;C12N1/20;C12N1/21;C12N5/00;C12N5/10;C12N15/00;C12N15/09;C12N15/19;C12N15/27;C12N15/70;C12N15/81;C12N15/85;C12P21/00;C12P21/02;C12R1/19;C12R1/865;C12R1/91;(IPC1-7):C12N15/27 |
主分类号 |
A61K35/74 |
代理机构 |
|
代理人 |
|
主权项 |
|
地址 |
|