Molecular cloning and expression of hybridoma growth factor in Escherichia coli.
Human monocytes produce a factor that supports the growth of B lymphocyte hybridoma cells, termed hybridoma growth factor (HGF). By using expression cloning in Escherichia coli of complementary DNA derived from human monocyte-poly(A+) RNA, we selected seven clones producing HGF activity as measured in a bioassay, based on the induction of proliferation of the HGF-dependent B cell hybridoma B9. Sequence analysis of the cDNA revealed that HGF is identical with interferon-beta 2, 26,000 protein, and B cell stimulatory factor-2. One of the active clones contained a cDNA that encoded a recombinant product lacking the 28-amino acid long signal peptide and the first 15 amino acids of the mature protein. Antibodies against the recombinant HGF inhibited the biologic activity of recombinant HGF as well as of monocyte-derived HGF.
Brakenhoff
J P
JP
Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam.
de Groot
E R
ER
Evers
R F
RF
Pannekoek
H
H
Aarden
L A
LA
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000187:drug effects; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000187:drug effects*; D007370:Interferon Type I / Q000235:genetics; D015850:Interleukin-6; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D007370:Interferon Type I; D015850:Interleukin-6; D007378:Interleukins; D011993:Recombinant Fusion Proteins; D004247:DNA