Related Experiment Video
Updated: Jun 17, 2026
Unraveling Key Players of Humoral Immunity: Advanced and Optimized Lymphocyte Isolation Protocol from Murine Peyer's Patches
Published on: November 21, 2018
[Effect of PTD-deltaPRD Foxp3 fusion protein on the mixed lymphocyte reaction in mice]
Weihong Shen1, Jing Liu, Xin Peng
1Department of Laboratory Medicine, Third People's Hospital of Wuxi, Wuxi Jiangsu 214041, China.
Objective:
To express protein transduction domain (PTD)-deletion proline domain (deltaPRD) Foxp3 fusion protein, and to analyze its influence on mixed lymphocyte reaction in mice.
Methods:
We cloned mouse deltaPRD of Foxp3 gene by PCR, and inserted it into pET28a-PTD, pET28a-PTD-eGFP vector, then expressed fusion proteins in E.coli Rosetta (DE3). The fusion proteins were purified and refolded by Profinity IMAC Ni(2+)-Charged Resin. The expression of fusion proteins was identified by Western blot. Flow cytometry assay was used to detect the effect of PTD-deltaPRD fusion protein to transduce into mouse EL-4 cells. The ability of fusion protein to inhibit the proliferation of EL-4 cells was analyzed by two-way mixed lymphocyte reaction.
Results:
The PTD-deltaPRD fusion proteins were expressed and purified efficiently. Western blot and flow cytometry indicated that PTD-deltaPRD fusion protein was transduced into EL-4 efficiently. Mixed lympocyte reaction assay showed that PTD-deltaPRD fusion protein had the bioactivity to inhibit the proliferation of EL-4 cells.
Conclusion:
The PTD-deltaPRD fusion protein was expressed in E.coli system and could be transduced into cells effectively, suggesting that PTD-deltaPRD fusion protein may be an inhibitor in lymphocytes from mouse spleen.
