DNA sequences encoding bovine interleukin-2
US4882282A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jul 31, 1986 |
| Grant date | Nov 21, 1989 |
| Priority date | — |
| Expiry date | Jul 31, 2006 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/75
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.