Cloning and expression analysis of Sirt2 in HEK293 cells / 细胞与分子免疫学杂志
Chinese Journal of Cellular and Molecular Immunology
;
(12): 970-972, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-622111
ABSTRACT
AIM:
To construct eukaryotic expression vector of Sirt2 and detect its expression in HEK293 cells.METHODS:
Total RNA was isolated from brain tissue of a-dult 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 se-quenced. 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.RESULTS:
The full length coding region of Sirt2 was obtained and confirmed by sequencing, the expression of Sirt2 was detected successfully in HEK293 cells.CONCLUSION:
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Cellular and Molecular Immunology
Year:
2009
Type:
Article
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