[Expression of recombinant human stem cell factor in insect cells].
The cDNA of human stem cell factor(hSCF) containing signal sequence was cloned into the transfer vector pVL941 of AcNPV to construct a recombinant transfer vector pVL941-SCF. Sf9 cells were cotransfected with wild type viral DNA and pVL914-SCF to produce the recombinant virus AcNPV-SCF by homologous recombination in cell. Southern-hybridization analysis suggested that the recombinant viral DNA contained hSCF cDNA fragment. The Sf9 cells infected with the recombinant baculovirus AcNPV-SCF expressed biologically active rhSCF which was secreted into the cell culture. The synergistic activities of SCF in conjunction with human interleukin-3(hIL-3) was measured by MTT colorimetric method and TF-1 cell line. The expression level of Sf9 cells reached its highest at about 1970 units/ml in the 3rd day after the infection with AcNPV-SCF. Three SCF bands with molecular masses of 18 x 10(3), 20 x 10(3) and 22 x 10(3) were detected by immunoblotting.
Zhang
D
D
Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093.
Qin
J
J
Zang
Y
Y
Zhu
J
J
Shen
B
B
D000818:Animals; D002478:Cells, Cultured; D006801:Humans; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D019089:Stem Cell Factor / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D011994:Recombinant Proteins; D019089:Stem Cell Factor