Purification and Refolding of LbSPL6 Recombinant Protein in Lycium barbarum
SPL(Squamosa promoter binding protein-like)is a plant-specific transcription factor that plays an important role in plant growth and development,secondary metabolism,and stress response.The SPL protein contains a highly conserved SBP functional domain,which regulates the expression of related genes by specifically binding to gene promoter elements.In this study,we optimized the induced expression conditions of the obtained Lycium barbarum pET28a-LbSPL6 fusion protein,and purified and refolded the formed inclusion bodies.The research results indicated that the expression of LbSPL6 protein was the highest under the conditions of an induction temperature of 37℃,IPTG final concentration of 1 mM,and an induction time of 4 hours.LbSPL6 protein exists in the form of inclusion bodies under various conditions.We recovered the inclusion bodies by ultrasonication,dissolved and renatured the precipitate in denaturation buffer,and purified the protein using Ni-NTA column affinity chromatography to obtain the active recombinant LbSPL6 fusion protein.The results of Western-blotting further confirmed that the purified protein was LbSPL6 recombinant protein.The purified recombinant LbSPL6 protein lays a foundation for further study on the function of Lycium barbarum LbSPL6 protein.
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