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- W2361307145 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 μg/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.
- W2361307145 created "2016-06-24" @default.
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- W2361307145 date "2002-01-01" @default.
- W2361307145 modified "2023-09-25" @default.
- W2361307145 title "Expression of Human Growth Hormone Gene in Bac-to-Bac Baculovirus System" @default.
- W2361307145 hasPublicationYear "2002" @default.
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