【摘要】 细胞自噬与植物生长、发育和逆境响应等过程密切相关。本研究构建了小麦重要自噬相关基因TaATG4a和TaATG8h的原核表达载体并导入大肠杆菌。IPTG诱导表达后的蛋白质SDS-PAGE电泳结果表明,两个基因在大肠杆菌中均能够高效表达,表达重组蛋白的分子量与预测分子量基本一致。利用Ni柱亲和层析方法进一步获得了纯度较高的重组蛋白,为今后的体外酶活分析、抗体制备和Western杂交等研究奠定了基础。更多还原

【Abstract】 Autophagy is closely related to the growth, development and stress responses of plant. In this study, prokaryotic expression vectors for wheat important autophagy-related gene TaATG4a and TaATG8h were constructed and transformed into E. coli. The results from SDS-PAGE showed that each gene could express efficiently in E. coli through IPTG induction. The molecular weight of each expressed recombinant protein was consistent with its predicted molecular weight. Recombinant proteins were further purified by the Ni-column affinity chromatography method. The obtaining of purified recombinant proteins laid a foundation for the future studies of in vitro enzyme activity, antibody preparation and western blot analysis.更多还原