AGL47 is a transcription factor coded by the At5g55690 gene on the fifth chromosome of Arabidopsis thaliana, belonging to MADS-box protein family. At5g55690 gene was reported to be upregulated by limited inorganic phosphate stress and may play an important role in plant phosphorus metabolism. To identify its function, At5g55690 gene' s full-length CDS was cloned to construct recombinant plasmid pET-28a-At5g55690. pET-28a-At5g55690 was then transformed into E. Coli BL21 ( DE3) cells to perform prokaryotic expression. AGL47 protein was obtained by induction of 1 mmol· L-1 IPTG but found to be in form of inclusion bodies. AGL47 soluble protein was isolated from purified inclusion bodies after denaturation with urea and renaturarion with dialysis.%AGL47是拟南芥第五染色体上AtSg55690基因编码的1个转录因子,属于MADS-box蛋白质家族,前期研究显示,At5g55690基因受磷胁迫特异诱导表达,极有可能在磷代谢调控中发挥重要的作用.为了进一步鉴定该基因的功能,本研究克隆At5g55690基因全长ORF,构建重组表达载体pPET-28a-At5g55690,转入表达宿主菌E.coli BL21( DE3),在1 mmol· L-1 IPTG诱导下,成功实现了AGL47蛋白的原核表达.可溶性鉴定结果显示,融合蛋白以包涵体的形式存在.对包涵体进行纯化,变性和复性后获得了可溶性目的蛋白.
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