Cloning and sequencing of cDNA encoding bovine macrophage colony-stimulating factor (bM-CSF) and expression of recombinant bM-CSF using baculovirus.
The cDNAs encoding bovine macrophage colony-stimulating factors alpha and beta (M-CSF alpha and M-CSF beta) were cloned and recombinant bovine M-CSF alpha (rbM-CSF beta) in its dimeric form was expressed by using a recombinant baculovirus/insect cell system. The predicted amino acid sequence of rbM-CSF alpha and rbM-CSF beta shared 83.3 and 75.9% (alpha), 75.3 and 65.9% (beta) similarity with the sequence for human and murine M-CSFs, respectively. The biological activity of rbM-CSF beta was confirmed by the colony-forming assay using mouse bone marrow cells. SDS-PAGE under a reducing condition showed that the molecular weight of rbM-CSF beta was approximately 34 kDa. On the other hand, Western blot analysis under a non-reducing condition revealed that this rbM-CSF beta was secreted in dimeric form into the cell supernatant.
Yoshihara
K
K
National Institute of Animal Health, Ibaraki, Japan. kazukazu@niah.affrc.go.jp
Inumaru
S
S
Hirota
Y
Y
Momotani
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D019281:Dimerization; D015870:Gene Expression; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D018411:Spodoptera
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D016173:Macrophage Colony-Stimulating Factor