The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Nov. 21, 1989

Filed:

Jul. 31, 1986
Applicant:
Inventors:

Dirk M Anderson, Seattle, WA (US);

Paul E Baker, Bainbridge Island, WA (US);

Michael A Cantrell, Seattle, WA (US);

Douglas P Cerretti, Seattle, WA (US);

David J Cosman, Seattle, WA (US);

Steven D Gimpel, Seattle, WA (US);

Kenneth H Grabstein, Seattle, WA (US);

Alf D Larsen, Seattle, WA (US);

Kate N McKereghan, Seattle, WA (US);

Assignee:

Immunex Corporation, Seattle, WA (US);

Attorneys:
Primary Examiner:
Assistant Examiner:
Int. Cl.
CPC ...
C12N / ; C12N / ; C12P / ; C12P / ;
U.S. Cl.
CPC ...
4352523 ; 435 68 ; 435 70 ; 435 91 ; 435255 ; 4351723 ; 435320 ; 536 27 ; 930 11 ; 930 29 ; 930 37 ; 930 38 ; 930 56 ; 930 60 ; 930 69 ;
Abstract

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.


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