Prokaryotic expression, purification and antigenicity identification of recombinant human survivin protein

• Published in: Xi bao yu fen zi mian yi xue za zhi Vol. 29; no. 8; p. 877
• Main Authors: Yin, Xiaotao, Wang, Wei, Tian, Renli, Xu, Yuanji, Yan, Jinqi, Zhang, Wei, Gao, Jiangping, Yu, Jiyun
• Format: Journal Article
• Language: Chinese
• Published: China 01.08.2013
• Subjects: Escherichia coli - genetics; Escherichia coli - metabolism; Genetic Vectors - genetics; Humans; Inhibitor of Apoptosis Proteins - biosynthesis; Inhibitor of Apoptosis Proteins - genetics; Inhibitor of Apoptosis Proteins - immunology; Inhibitor of Apoptosis Proteins - isolation & purification; Plasmids - genetics; Prokaryotic Cells - chemistry; Prokaryotic Cells - metabolism; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - isolation & purification
• ISSN: 1007-8738

To construct a prokaryotic expression plasmid pET28a-survivin, optimize the recombinant protein expression conditions in E.coli, and purify the survivin recombinant protein and identify its antigenicity. Survivin cDNA segment was amplified by PCR and cloned into prokaryotic expression vector pET28a(+) to construct the recombinant expression vector pET28a-survivin. The expression vector was transformed into BL21 (DE3) and the fusion protein survivin/His was induced by IPTG. The fusion protein was purified through Ni affinity chromatography. The antigenicity of the purified survivin protein was identified by Western blotting and ELISA. The recombinant expression vector was verified successfully by BamHI and HindIII. The fusion protein induced by IPTG was obtained with Mr; about 24 000. The purity of the purified protein reached 90% by SDS-PAGE analysis. And the antigenicity of the survivin protein was validated by Western blotting and ELISA. The prokaryotic expression plasmid pET28a-survivin was successfully co