Molecular Cloning of a Complementary DNA Encoding Human Macrophage-Specific Colony-Stimulating Factor (CSF-1)

Abstract

Complementary DNA (cDNA) clones encoding human macrophage-specific specific colony-stimulating factor (CSF-1) were isolated. One cDNA clone codes for a mature polypeptide of 224 amino acids and a putative leader of 32 amino acids. This cDNA, which was cloned in the Okayama-Berg expression vector, specifies the synthesis of biologically active CSF-1 in COS cells, as determined by a specific radioreceptor assay, macrophage bone marrow colony formation, and antibody neutralization. Most of the cDNA isolates contain part of an intron sequence that changes the reading frame, resulting in an abrupt termination of translation; these cDNA's were inactive in COS cells. The CSF-1 appears to be encoded by a single-copy gene, but its expression results in the synthesis of several messenger RNA species, ranging in size from about 1.5 to 4.5 kilobases.

Macrophage colony‐stimulating factor purified from normal human urine Amino‐terminal sequence and amino acid composition

“…The sequence deduced by Kawasaki et al [2] is nearly consistent with that deduced by Wong et al [3] between position 1 and position 148. However, the latter sequence contains an additional sequence of 295 residues intervening between position 148 and position 149 of the former.…”

“…The analytical values are expressed as mol individual amino acid per mol His, and the number of residues of each amino acid was calculated with respect to the integral value of His being 4 or 5. Table 2 also shows the number of amino acid residues present in the sequence deduced by other workers [2,3].…”

“…Kawasaki et al [2] also determined the Nterminal 12 residues of human urine M-CSF and the N-terminal 30 residues of mouse L-cell M-CSF. The sequence obtained by protein sequencing exactly matched that deduced from the analysis of cDNA insofar as the first 12 residues of human M-CSF were concerned.…”

“…The total amino acid sequence of human M-CSF was deduced by Kawasaki et al [2] as well as by Wong et al [3] from the nucleotide sequence of complementary DNA (cDNA) clones derived from human tumor cell lines. The sequence deduced by Kawasaki et al [2] is nearly consistent with that deduced by Wong et al [3] between position 1 and position 148.…”

Modulation of c-fms proto-oncogene in an ovarian carcinoma cell line by a hammerhead ribozyme

“…The length of the PCR product was 406 bases. The primers for M-CSF and G3PDH were derived from the work of other investigators (Kawasaki et al, 1985;Adcock et al, 1994) (M-CSF: upstream primer, 5'-ACGACATGGCTGGGCTCCCT and downstream primer, 5'-TTCTCCAGCAACTGGAGAGGTG. G3PDH: upstream primer, 5'-TGAAGGTCGGAGTCAACGGAThTTGGT and downstream primer, 5'-CATGTGGGCCATGAGGTCCAC-CAC).…”