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- W3150327846 abstract "The Bac-to-Bac baculovirus expression system is a novel gene expression system that allows the rapid and efficient generation of recombinant baculovirus DNA by site-specific transposition in E. coli, rather than homologous recombination in insect cells. The cDNA encoding hGH (human growth hormone) gene was cloned into a transfer vector pFastBac1, and recombinant vector pFastBac-hGH DNA was extracted. After transformation, it was introduced into E. coli DH10Bac which included a shuttle vector, Bacmid. By site-specific transposition, hGH gene was integrated into Bacmid, and a recombinant shuttle vector was constructed, named rBacmid-hGH. The cultured Sf9 cells were directly transfected with the rBacmid-hGH DNA, and the pure recombinant baculovirus was obtained, named rAcV-Bac-hGH. RE assay, PCR and Southern blot analysis revealed that hGH cDNA was correctly inserted into baculovirus genome under the control of polyhedrin promoter. The hGH expression product had a Mr of 22 kD. Immuno-Chemiluminescent was used for detecting hGH expression level. The hGH expression level in transfection supernatant was 18 #mu#g/ml, 400 times more than that in the co-transfection supernatant by the traditional BEVS (40 ng/ml). Advantages of using this system in the expression control of hGH gene in vitro were also discussed." @default.
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- W3150327846 date "2002-01-01" @default.
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- W3150327846 title "Expression of human growth hormone gene in Bac-to-Bac baculovirus system" @default.
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