Expression, purification, and activity of sika deer prothymosin α protein
Dongming Xue1, Liyue Zhou1, Tianxia Sun1
1Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, 130117, PR China.
Objective:
To perform the prokaryotic expression and purification of sika deer prothymosin α (PTMA) protein and lay a foundation for the subsequent study of PTMA protein activity.
Methods:
The PTMA fragment was synthesized and cloned into the pET21a-PTMA vector. Escherichia coli BL21 (DE3) was used for the prokaryotic expression of the PTMA protein, and the expression products were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nickel-containing electrophoresis. Polyacrylamide gel electrophoresis was conducted to analyze the expression products, which were purified using nickel ion affinity chromatography. Fragments were identified using mass spectrometry and analyzed for activity in MC3T3-E1 and ATDC5 cells.
Results:
The pET21a-PTMA expression vector was successfully constructed, and the protein purity reached more than 93 %. The purified protein displayed proliferative activity in both MC3T3-E1 and ATDC5 cells.
Conclusion:
The successful construction of a stable expression vector and production of high-purity protein lay a foundation for future research on PTMA activity and its effects on bone diseases.
