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Updated: Aug 16, 2026
Purification of Ubiquitinated p53 Proteins from Mammalian Cells
Published on: March 21, 2022
[Expression, purification and transduction of Tat-p53 fusion protein]
Jin Ding1, Jun Liu, Yu-xiao Huang
1Department of Etiology, Fourth Military Medical University, Xi'an 710032, China. dingjin1103@sina.com
Aim:
To express and purify Tat-p53 fusion protein and investigate its transduction efficiency.
Methods:
The gene encoding wide-type p53 was isolated using RT-PCR from A549 cell line and cloned into pTAT-HA and pET32a prokaryotic expression vectors. Recombinant plasmids were transformed into E.coli BL21(DE3)LysS, then the transformed cells were induced with IPTG. The expression and purification of the Tat-p53 and p53 were analyzed by SDS-PAGE. BALB/c mice were immunized with purified p53 protein. The serum was isolated and the antibody specific to p53 was measured by ELISA. The transduction efficiency of Tat-p53 was detected using indirect immunofluorescence assay.
Results:
Prokaryotic expression vectors of Tat-p53 and p53 were constructed correctly. Tat-p53 fusion protein and p53 protein were successfully expressed and purified. p53 specific mouse antiserum was obtained. IFA result indicated that Tat-p53 fusion protein transduced into HepG2 cells efficiently.
Conclusion:
The obtained Tat-p53 fusion protein may be valuable for the basic research on therapy for liver carcinoma.
