PEP－1－SODl融合蛋白的制备、表达及纯化

PEP-1-SOD1 fusion protein：preparation, expression and purification

目的：构建原核表达载体pQE30－PEP－1－SOD1，并进行PEP－1－SOD1融合蛋白表达和纯化。方法载体质粒pQE－30及模板质粒pUC57－PEP－1－SOD1分别进行酶切后，构建原核表达载体pQE30－PEP－1－SOD1载体。经测序证实构建成功后，转化JM109，表达PEP－1－SOD1融合蛋白，并进行鉴定。结果 SDS－PAGE电泳分析表明，位于相对分子质量约25×10^3处出现PEP－1－SOD1的目标表达条带；目的蛋白以天然的可溶性的形式存在。结论已成功制备出PEP－1－SOD1融合蛋白。

Objective To establish a prokaryotic expression vector pQE 30-PEP-1-SOD1 and express and purify PEP-1-SOD1 fusion protein .Methods The vector pQE30-PEP-1-SOD1 was established using a plasmid pQE -30 and a template plasmid pUC57-PEP-1-SOD1 through enzymatic digestion .The recombinant plasmid was then se-quenced before transformed into JM109 for expression of PEP -1-SOD1 fusion protein.The resultant products were i-dentified by SDS-PAPG electrophoresis.Results According to SDS-PAGE analysis, the target protein was present in a soluble form with about 25×10 3 molecular weight .Conclusion The fusion protein PEP -1-SOD1 is successfully prepared.