农业科学研究2024，Vol.45Issue(3)：49-54,6.
枸杞LbSPL6重组蛋白的纯化和复性
Purification and Refolding of LbSPL6 Recombinant Protein in Lycium barbarum
摘要
Abstract
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.关键词
枸杞/LbSPL6重组蛋白/蛋白质纯化/蛋白质复性Key words
Lycium barbarum/LbSPL6/protein purification/protein refolding分类
农业科技引用本文复制引用
张金金,张真露,石晶..枸杞LbSPL6重组蛋白的纯化和复性[J].农业科学研究,2024,45(3):49-54,6.基金项目
宁夏自然科学基金项目(2023AAC03123) （2023AAC03123）
中央引导地方科技发展资金项目(2023FRD05032) （2023FRD05032）
宁夏回族自治区重点研发计划重点项目(2022BBF02010) （2022BBF02010）
