PURPOSE: To enable obtaining of a pharmaceutically useful substance in a large amount by containing a specific base sequence in a gene and making the gene code a polypeptide having a monocytemacrophage colony stimulating factor (N-CSF) activity.
CONSTITUTION: An M-CSF-producing human monocytic cell strain U937 is cultured in a culture medium containing phorbol myristate acetate and cycloheximide which are immunological stimulating substances to provide cultured microbial cells (A), which are then extracted to afford an extract. The resultant extract is subsequently fractionated by a sucrose density gradient centrifugation method to afford an mRNA (B) having a value of 24.7S to 27.0S and polyadenylic acid structure at the 3'-terminal. cDNA is then prepared from the aforementioned ingredient (B) to provide a genetic library, which is subsequently selected with a desired probe to produce a gene (C) capable of coding a polypeptide containing an amino acid sequence expressed by formula I, etc., and and coding the N-CSF active polypeptide composed of a base sequence expressed by formula II, etc. A transformant strain containing, as necessary, the ingredient (C) is cultured to produce the M-CSF-active substance.
| JP5756407 | Backflow purification of polypeptides |
| JP4671372 | Purification of neurotrophins |
| WO/2012/030512 | FLOW-THROUGH PROTEIN PURIFICATION PROCESS |
OGAWA HIROYUKI
SAKAI NOBUO
MIMURA SHUJI
SUZUKI HIROYASU
| JPH01104176A | 1989-04-21 | |||
| JPH022391A | 1990-01-08 | |||
| JPS6422899A | 1989-01-25 |
| WO1988008003A1 | 1988-10-20 | |||
| WO1988003173A2 | 1988-05-05 |