发明授权
- 专利标题: DNA sequences encoding bovine interleukin-2
- 专利标题(中): 编码牛白介素-2的DNA序列
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申请号: US888994申请日: 1986-07-31
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公开(公告)号: US4882282A公开(公告)日: 1989-11-21
- 发明人: Dirk M. Anderson , Paul E. Baker , Michael A. Cantrell , Douglas P. Cerretti , David J. Cosman , Steven D. Gimpel , Kenneth H. Grabstein , Alf D. Larsen , Kate N. McKereghan
- 申请人: Dirk M. Anderson , Paul E. Baker , Michael A. Cantrell , Douglas P. Cerretti , David J. Cosman , Steven D. Gimpel , Kenneth H. Grabstein , Alf D. Larsen , Kate N. McKereghan
- 申请人地址: WA Seattle
- 专利权人: Immunex Corporation
- 当前专利权人: Immunex Corporation
- 当前专利权人地址: WA Seattle
- 主分类号: C12N15/09
- IPC分类号: C12N15/09 ; A61K35/12 ; A61K35/14 ; A61K35/74 ; A61K38/00 ; C07H21/04 ; C07K1/20 ; C07K14/00 ; C07K14/52 ; C07K14/54 ; C07K14/55 ; C12N1/00 ; C12N1/19 ; C12N1/21 ; C12N15/00 ; C12N15/73 ; C12P21/02 ; C12R1/19 ; C12R1/865
摘要:
A chemically-synthesized oligonucleotide composing a portion of the nucleotide sequence of the human IL-2 is employed as a probe to isolate the gene coding for human IL-2. The human IL-2 gene is selected from a cDNA library prepared from RNA produced by mitogen-stimulated Jurkat cells. Double-stranded cDNA is prepared from polyadenylated RNA extracted from bovine cells thought to produce interleukin-2. Such cDNA is inserted within a plasmid vector and the recombinant plasmid employed to transform hosts. Plasmid DNA, prepared from pools of the transformed hosts, is hybridized with a probe composed of a large portion of the coding sequence of the human IL-2 gene. Pools of host cells that provide signal to the human cDNA probe are identified, subdivided, and rescreened until a single positive colony is identified. Bovine plasmid cDNA is prepared from this colony, and the bIL-2 gene is sequenced. The plasmid DNA is employed to express recombinant bovine IL-2 in yeast and bacterial expression systems, with the expressed bovine IL-2 purified to homogeneity by one or more reverse phase high-performance liquid chromatography procedures.
公开/授权文献
- US5887607A Wafer processing apparatus 公开/授权日:1999-03-30
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