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Yeast‐prepro‐alpha‐factor‐leader‐region‐directed synthesis and secretion of truncated human macrophage colony‐stimulating factor in the silkworm Bombyx mori
Author(s) -
Qiu P.,
Qin J.,
Ding Y.,
Zhu D.
Publication year - 1995
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1995.tb00326.x
Subject(s) - bombyx mori , biology , bombycidae , signal peptide , hemolymph , microbiology and biotechnology , recombinant dna , yeast , nuclear polyhedrosis virus , biochemistry , bombyx , gene
Human macrophage colony‐stimulating factor (hM‐CSF) cDNA joined to the leader region of the precursor of the yeast mating pheromone alpha‐factor (MF alpha L) was expressed at high levels in BmN cells and in silkworm (Bombyx mori) larvae, using recombinant Bombyx mori nuclear polyhedrosis virus, as a vector. The biological activity of rhM‐CSF detected in the haemolymph was 1 times 10(6) colony‐formation units/ml, approximately half of the expression level directed by the native signal peptide of hM‐CSF in silkworm larvae. The secreted rhM‐CSF was purified to homogeneity. N‐terminal analysis showed that the signal peptide had been removed, indicating that insect cells possess the enzymic activity necessary to cleave the pro‐alpha‐factor leader region from the fusion protein at the carboxy side of Lys‐Arg dibasic residues, which is the cleavage site recognized by KEX2 endopeptidase in yeast cells.