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[Cloning and expression analysis of Sirt2 in HEK293 cells].

Authors: Tao, Wang; Wei, Wang; Qing, Xu; Lu-yuan, Guan; Bin, Zhang;

[Cloning and expression analysis of Sirt2 in HEK293 cells].

Abstract

To construct eukaryotic expression vector of Sirt2 and detect its expression in HEK293 cells.Total RNA was isolated from brain tissue of adult SD rat. A 1 130 bp fragment containing the coding region of Sirt2 was amplified by RT-PCR and the resulting PCR product was subcloned into PMD20-T vector and sequenced. Coding region of Sirt2 was generated with PCR by using the PMD20-T-Sirt2 as template, the amplified PCR fragment was inserted into the EcoR I and Hind III sites of the pcDNA3.1myc-his(-)A expression vector, and the sequence was confirmed by DNA sequencing. The expression of new construct pcDNA3.1 myc-his(-)A-Sirt2 in HEK293 cells was detected by immunofluorescence.The full length coding region of Sirt2 was obtained and confirmed by sequencing, the expression of Sirt2 was detected successfully in HEK293 cells.The eukaryotic expression vector of Sirt2 has been successfully constructed, which will provide a useful tool for designing an in-depth investigation of the role of Sirt2.

Related Organizations
Keywords

Male, DNA, Complementary, Sirtuin 2, Genetic Vectors, Animals, Gene Expression, Humans, Cloning, Molecular, Cell Line, Plasmids, Rats

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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Average
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