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- W2388350209 abstract "[Objective] The expression of rat DNA polymerase β was up-regulated after transient middle cerebral artery occlusion, using the technique of fluorescence differential display reverse transcription polymerase chain reaction (FDD RT-PCR). To determine the role of DNA polymerase β (polβ) in brain injury. An adenoviral vector was constructed containing the coding sequence for rat DNA polymerase β. [Methods] The rat po1β coding sequenee was cloned to pGEM-T Easy~(TM) vector by RT-PCR, and confirmed by DNA sequencing and subcloned into shuttle plasmid pAdTrack-CMV, then linearized and transferred to E. coli (BJ5183) contained adenoviral backbone vector pAdEasy-1. The recombinant plasmid was packaged and amplified in HEK 293 cells. PC12 cells were infected with the recombinant adenoviruses. [Results] Screening 10 pGEM-T Easy~(TM) clones, 3 clones have no base changes. Eight out of 30 homologous recombination plasmids have the target band. The Adpolβb recombinant plasmid was verified by DNA sequence analysis. After infected with Adpolβ adenovirus, strong green fluorescence were observed in HEK 293 and PC12 cells under fluorescence microscopy. Adpolβ DNA in 293 cells was identified by PCR. The expression of rat DNA polymerase β in Adpolβ infected PC12 cells are eonfirmed by immunoblot analysis. [Conclusion] Recombinant adenoviral vectors containing rat DNA polymerase β were constructed and efficiently expressed in PC12 cells." @default.
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- W2388350209 date "2005-01-01" @default.
- W2388350209 modified "2023-09-23" @default.
- W2388350209 title "Construction of Recombinant Adenovirus Containing Rat DNA Polymerase β and Its Expression in PC12 Cells" @default.
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