[Effect of PTD-deltaPRD Foxp3 fusion protein on the mixed lymphocyte reaction in mice].
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.


CONCLUSIONS
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.
Shen
Weihong
W
Department of Laboratory Medicine, Third People's Hospital of Wuxi, Wuxi Jiangsu 214041, China.
Liu
Jing
J
Peng
Xin
X
Zong
Yangyong
Y
Xu
Xun
X
Shao
Qixiang
Q
D000818:Animals; D049109:Cell Proliferation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D051858:Forkhead Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D017353:Gene Deletion; D005822:Genetic Vectors / Q000235:genetics; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D011392:Proline / Q000235:genetics* / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology
D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D011993:Recombinant Fusion Proteins; D011392:Proline