Expression of granulocyte colony-stimulating factor by human cell lines.
We have isolated and expressed a cDNA clone that encodes a human granulocyte colony-stimulating factor from the MIA PaCa-2 cell line. A genomic clone of this factor has been isolated from the CHU-2 cell line and is reported to encode two alternative transcripts [The EMBO J. 5,575, 1986]; one transcript predicts an amino acid sequence identical to that predicted by our MIA PaCa-2 cDNA clone; the other transcript predicts a similar protein containing a three amino acid residue insertion. To investigate which types of this colony-stimulating factor are produced by other cell lines, we used specific oligonucleotides to determine which types of transcripts were present in MIA PaCa-2, 5637, and LD-1 cells, all of which have been reported to produce a factor that can stimulate the growth of predominantly granulocyte colonies in human bone marrow cell cultures. Northern analysis with these probes revealed MIA PaCa-2-like transcripts in all of these cell lines and failed to detect transcripts that would encode the colony-stimulating factor that contained the three-amino-acid-residue insertion.
D001483:Base Sequence; D001854:Bone Marrow Cells; D002460:Cell Line; D003101:Collodion; D004247:DNA / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics* / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000378:metabolism; D014158:Transcription, Genetic
D007377:Interleukin-3; D009841:Oligonucleotides; D003101:Collodion; D004247:DNA