DNA Encoding human colony stimulating factor, peptide encoded thereby,vectors and transformed hosts containing such DNA, and the production of all thereof

Abstract

Abstract A chemically-synthesized oligonucleotide composing a portion of the nucleotide sequence of the murine GM-CSF may be employed as a probe to isolate the gene coding for murine GM-CSF. The murine GM-CSF gene may be selected from a murine cDNA library prepared from RNA produced by mitogen-stimulated LBRM-33 splenic lymphoma cells. Double-stranded cDNA may be prepared from polyadenylated RNA extracted from human cells thought to produce human CM-CSF. Such cDNA is insertable within a plasmid vector and the recombinant plasmid employed to transform hosts. Plasmid DNA, prepared from pools of the transformed hosts, may be hybridized with a probe composed of a large portion of the coding sequence of the murine GM-CSF cDNA gene. Pools of host cells that provide a positive signal to the murine cDNA probe may be identified, subdivided, and rescreened until a single positive colony is identified. Human plasmid cDNA can be prepared from this colony, and the human GM-CSF gene sequenced. The coding region of the gene may be inserted into an expression vector for expression of functional human GM-CSF.

Inventors

• 👤 Dirk M Anderson
• 👤 Michael A Cantrell
• 👤 Douglas P Cerretti
• 👤 David J Cosman
• 👤 Alf D Larsen
• 👤 Virginia L Price

Bibliographic Details