A novel recombinant dual human SCF expressed in and purified from silkworm, Bombyx mori, possesses higher bioactivity than recombinant monomeric human SCF.
A novel recombinant dual human stem cell factor (rdhSCF) gene was constructed which consisted of a full-length hSCF cDNA plus a truncated hSCF cDNA (1-145 aa), linked by a peptide (GGGGSGGGGSGG) coding region. The rdhSCF gene was cloned into baculovirus transfer vector pAcSecG2T under the polyhedrin promoter control. Silkworm larvae infected with the recombinant virus expressed rdhSCF up to 15,800 units/mL in haemolymph. The specific activity of rdhSCF purified from the haemolymph was up to 3.0 x 10(6) units/mg, about 8.6 times as high as that of monomer rhSCF from Escherichia coli, and about 9.1 times as high as that of monomer rhSCF from insect cell. The binding affinity of rdhSCF to the cell surface receptor was higher than that of monomer rhSCF.
Han
Junhai
J
School of Life Science and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
Zang
Yuhui
Y
Lu
Haiqin
H
Zhu
Jie
J
Qin
Junchuan
J
D000818:Animals; D012831:Bombyx; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019281:Dimerization; D004926:Escherichia coli; D005813:Genes, Synthetic; D005822:Genetic Vectors / Q000235:genetics; D006458:Hemolymph / Q000737:chemistry; D006801:Humans; D007814:Larva; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011485:Protein Binding; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology; D011995:Recombination, Genetic; D018411:Spodoptera; D019089:Stem Cell Factor / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D019089:Stem Cell Factor; D019009:Proto-Oncogene Proteins c-kit