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Updated: Jun 28, 2026

Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
Published on: March 10, 2023
[Construction of different mutants of HA-tagged human RAGE gene and their eukaryotic expression]
Wei-wei Cheng1, Yu-sheng Li, Xiao-wei Gong
1Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China. chengweiwei1984@163.com
Objective:
To construct eukaryotic expression vectors for HA-tagged receptor for advanced glycation end products (RAGE) mutants.
Methods:
Site-directed mutagenesis was applied to wild-type RAGE gene cloned in the pcDNA3 vector with HA tag to obtain the mutants pcDNA3-HA-RAGE(S391A), pcDNA3-HA-RAGE(S399A), pcDNA3-HA-RAGE(S400A), and pcDNA3-HA-RAGE(T401A). After identification by sequencing, the mutants were transfected into HEK293 cells, and the expression of these mutants were detected by Western blotting using anti-HA antibody.
Results:
The HA-tagged RAGE mutants constructed were verified successfully by sequencing, and highly expressed in HEK293 cells.
Conclusion:
The success in constructing HA-tagged RAGE mutants, which are highly expressed in eukaryotic cells, may facilitate the functional study of RAGE in cell signal transduction.
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