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- W2372695784 abstract "Enhanced green fluorescent protein(EGFP), myc epitope and polyhistidine metalbinding tag are often used as a marker for recombinant fusion protein in many gene expression vectors, each marker has its own function, EGFP emits green fluorescence for direct detection, myc epitope facilitates recombinant fusion protein detection using its antibodies, polyhistidine tag allows purification of recombinant fusion protein using resin. Hitherto, no a plasmid vector can integrate!all of these functions. In this study we constructed a novel eukaryotic expressive plasmid, designated as pcDNA6/mychisEGFP B, which integrated the functions of EGFP, myc epitope and polyhistidine tag. Importantly, a linker octo-peptide in N terminal of EGFP was designed using LINKER program. A DNA fragment encoding a putative protein containing a signal peptide of human interleukin 2(IL2) in N terminal was cloned into pcDNA6/mychisEGFP B in frame with the Cterminal peptide to construct pMHES. 2.2.15 Cells were transfected with pcDNA6/mychisEGFP B and pMHES, and Balb/c mice were intravenously injected with pcDNA6/mychisEGFP B by tails, results revealed that both of the plasmids worked in 2.2.15 Cells and livers of Balb/c mice. Assuming gene of the IL2 was inserted into pcDNA6/mychisEGFP B in frame with EGFP, myc and 6×His, threedimensional structure for this putative expression product was simulated using Modeller8V2, results revealed that IL2, EGFP, myc and 6×his did not interfere each other and octo-peptide linker owned certain flexibility. The results suggest that pcDNA6/mychisEGFP B may be useful as a genetic tool for mammalian cells and a vector for gene therapy." @default.
- W2372695784 created "2016-06-24" @default.
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- W2372695784 date "2006-01-01" @default.
- W2372695784 modified "2023-09-25" @default.
- W2372695784 title "Construction of a Novel Eukaryotic Expression Plasmid pcDNA6/myc-his-EGFP Band Its Applications in Expression of Recombinant Genes" @default.
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