| 盖文丽颜冬菁王伟陈孝平陈正望△.生物活性肽Lunasin 的原核表达和分离纯化[J].,2011,11(5):805-807 |
| 生物活性肽Lunasin 的原核表达和分离纯化 |
| Prokaryotic Expression and Purification of Bioactive Peptide Lunasin |
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| DOI： |
| 中文关键词: 原核表达 亲和层析 IPTG |
| 英文关键词: Prokaryotic Expression Affinity Chromatography IPTG |
| 基金项目:国家自然科学基金（30470823） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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). |
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