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分析大豆Gm NF-YA3基因结构,构建Gm NF-YA3基因的原核表达载体并进行表达,以期获得带有GST标签的目的蛋白。通过Phytozome数据库对Gm NF-YA3基因组DNA序列进行生物信息学分析,序列比对结果表明,Gm NF-YA3基因组DNA序列长度为4 589bp,由5个外显子和4个内含子组成。为了获得纯化的Gm NF-YA3蛋白,将保守域片段亚克隆到原核表达载体p GEX4T-1中,SDS-PAGE表明融合蛋白受异丙基硫代半乳糖苷(IPTG)诱导高水平表达,但主要以包涵体形式存在。本研究为进一步纯化和鉴定Gm NF-YA3蛋白及研究其功能奠定了基础。
The gene structure of Gm NF-YA3 was analyzed and the prokaryotic expression vector of Gm NF-YA3 gene was constructed and expressed in order to obtain the target protein with GST tag. The bioinformatics analysis of Gm NF-YA3 genomic DNA sequences by Phytozome database showed that the length of Gm NF-YA3 genomic DNA was 4 589bp and consisted of 5 exons and 4 introns. In order to obtain the purified Gm NF-YA3 protein, the conserved domain was subcloned into the prokaryotic expression vector pGEX4T-1. SDS-PAGE showed that the fusion protein was induced by IPTG at a high level, Mainly in the form of inclusion. This study laid the foundation for further purification and identification of Gm NF-YA3 protein and its function.