生物活性肽Lunasin的原核表达和分离纯化

Prokaryotic Expression and Purification of Bioactive Peptide Lunasin

目的:利用基因工程的方法在大肠杆菌中表达并纯化生物活性肽Lunasin。方法:将合成的Lunasin基因插入原核表达载体pET-32a(+)的多克隆位点Nde I和Xho I之间,然后将重组载体转化入大肠杆菌BL21(DE3)中,利用IPTG诱导表达蛋白,经SDS-PAGE和Western Blot鉴定蛋白的表达。然后利用亲和层析技术将含有6×His标签的蛋白分离纯化、脱盐、冻干。结果:①鉴定结果表明在6kDa位置出现目的条带Lunasin重组蛋白。②亲和层析在100mM咪唑时得到了洗脱的重组蛋白。结论:在大肠杆菌BL21(DE3)中成功表达并且纯化出了生物活性肽Lunasin。

Objective：To express prokaryotic and purify bioactive peptide Lunasin in E.coli by genetic engineering methods.Methods：The gene of Lunasin was artificially synthetic and then inserted into Nde I and Xho I enzyme-cutting sites of pET-32a（＋） plasmid.The recombinant expression vector pET-32a（＋）-Lunasin-6His was transformed into E.coli BL21（DE3）.The fusion protein Lunasin was solubly expressed after the induction of IPTG.The fusion protein was identified by SDS-PAGE and Western Blot.After the expressed protein was purified by affinity binding chromatography with Ni-NTA,salt-out and freeze-dried.Results：① The molecular mass of Lunasin-6His was 6kDa.② The fusion protein was eluted by 100 mM Imidazole.Conclusion：Bioactive peptide Lunasin was successfully expressed and purified in E.coli BL21（DE3）.