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- W2376874625 abstract "Objective To construct a recombinant adenovirus vector for tissue kallikrein 1(TK1) and tissue inhibitor of matrix metalloproteinase(TIMP) gene expression,so as to investigate the synergistic effects of TKl combination with TEMPI on improving vascular restenosis.Methods The human TIMPI gene in original plasmid was synthesized and digested with endonuclease Not I /Hind III,and cloned to shutde plasmid pDC316with mCMV promoter to construct recombinant shutde plasmid.The mCMV-hTIMPl sequences in the shuttle plasmid was then digested with endonuclease BglII/SalI and inserted into the plasmid pDC316-hTKl(saved in our institute) to construct recombinant shuttle plasmid(with double gene hTK1 and hTIMP1).The shuttle plasmids and adenovirus backbones pBHGlox_El,3Cre in 293A cells were recombined by AdMax with Cre/loxP,and packaged with the help of Iipofectamine 2000,respectively.The expression of hTKland hTIMPl proteins was confirmed by Western blot analysis.Results The two recombinant shutde plasmids pDC316-hTIMPl and pDC316-hTKl-hTIMPl were constructed successfully as proved by the methods of PCR,restriction and sequencing analysis.Recombinant adenoviral vectors Ad-hTKl-hTIMPl was successfully packaged in 293A cells.The abundance expression of hTKland hTIMPl proteins were detected in VSMCs transfected with Ad-hTKl-hTIMPl,respectively and independently.Conclusion A recombinant adenovirus vector containing hTK1 and hTIMP1 gene with independent promoter was successfully constructed and co-expressed in rat VSMCs,which laid a foundation for the gene therapy study on vascular remodeling diseases." @default.
- W2376874625 created "2016-06-24" @default.
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- W2376874625 date "2013-01-01" @default.
- W2376874625 modified "2023-09-26" @default.
- W2376874625 title "Construction of Recombinant Adenovirus Vector Expressing TK1 and TIMP1 Genes and Co-expression in Rat Vascular Smooth Muscle Cells" @default.
- W2376874625 hasPublicationYear "2013" @default.
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