| Title: | 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; |
| Address: | "Department of Biochemistry, Nanjing University, People's Republic of China" |
| ISSN/ISBN: | 0885-4513 (Print) 0885-4513 (Linking) |
| Abstract: | "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 x 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" |
| Keywords: | "Amino Acid Sequence Animals Base Sequence Bombyx/genetics/*metabolism Cells, Cultured DNA/chemistry/genetics Electrophoresis, Polyacrylamide Gel Gene Expression Regulation, Bacterial/genetics Genetic Vectors Humans In Situ Hybridization Larva/cytology/met;" |
| Notes: | "MedlineQiu, P Qin, J Ding, Y Zhu, D eng Research Support, Non-U.S. Gov't 1995/02/01 Biotechnol Appl Biochem. 1995 Feb; 21(1):67-75" |
