Abstract:

Objective To study cloning, prokaryotic expression and purification of human synuclein gamma.
Methods A DNA fragment encoding human synuclein gamma was amplified by polymerase chain reaction from a human cDNA library and was cloned into vector pGEX-6p-1. The construct carrying the coding DNA sequence of synuclein gamma fused with GST was transformed into E.coli BL21(DE3), and the fusion protein of synuclein gamma and GST was expressed by induction with IPTG. The product was purified by affinity chromatography, ion exchange chromatography and gel filtration.
Results Human synuclein gamma was successfully expressed in E.coli in soluble form. The product was identified to be human synuclein gamma by SDS-PAGE and mass spectrography. After purification, the purity of the recombinant protein was more than 98%.
Conclusion An effective purification protocol was established. Recombinant synuclein gamma with a purity more than 98% was obtained, which provided a basis for the further studies of the crystal structure, biological activities and functions of synuclein gamma.

Key words: synuclein gamma, fusion protein, prokaryotic expression, purification