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Sirt1 Gene Eukaryotic Eexpression of SD Rat

Lan Hu

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Abstract

Sirt1 cDNA containing XhoI/EcoRI sites was amplified by RT-PCR,and then cloned and sequenced.Eukaryotic ex-pression vector pEGFP-N1-Sirt1 was transfected into 293 cells with lipidosome,and the expression protein was detected by SDS-PAGE.The results showed that pEGFP-N1-Sirt1 could be transfected into 293 cells with lipidosome successfully.The green flu-orescence protein was observed 48h after transfection.The fusion protein was detected with SDS-PAGE,which was about 41kD.The results provided experimental basis for further study on the function of Sirt1 gene.

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What this paper is about

Sirt1 cDNA containing XhoI/EcoRI sites was amplified by RT-PCR,and then cloned and sequenced.Eukaryotic ex-pression vector pEGFP-N1-Sirt1 was transfected into 293 cells with lipidosome,and the expression protein was detected by SDS-PAGE.The results showed that pEGFP-N1-Sirt1 could be transfected into 293 cells with lipidosome successfully.The green flu-orescence protein was observed 48h after transfection.The fusion protein was detected with SDS-PAGE,which was about 41kD.The results provided experimental basis for further study on the function of Sirt1 gene.

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Available abstract

Sirt1 cDNA containing XhoI/EcoRI sites was amplified by RT-PCR,and then cloned and sequenced.Eukaryotic ex-pression vector pEGFP-N1-Sirt1 was transfected into 293 cells with lipidosome,and the expression protein was detected by SDS-PAGE.The results showed that pEGFP-N1-Sirt1 could be transfected into 293 cells with lipidosome successfully.The green flu-orescence protein was observed 48h after transfection.The fusion protein was detected with SDS-PAGE,which was about 41kD.The results provided experimental basis for further study on the function of Sirt1 gene.

Key concepts: XhoI, Transfection, Molecular biology, EcoRI, Complementary DNA, Fusion protein, Gene, Biology

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