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Expression of Human Stem Cell Factor with Recombinant Baculovirus in BmN Cell Line and Silkworm  

Xijie, Guo (Sericultural Research Institute, Chinese Academy of Agricultural Sciences)
Yongfeng, Jin (Institute of Biochemistry, Chinese Academy of Agricultural Sciences)
Mingguan, Yang (College of Animal Sciences, Zhejiang University)
Yaozhou, Zhang (Institute of Biochemistry, Chinese Academy of Agricultural Sciences)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.4, no.1, 2002 , pp. 51-56 More about this Journal
Abstract
A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.
Keywords
Human stem cell factor; Baculovirus vector expression system; Expression;
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