布鲁菌外膜蛋白Bp26在大肠埃希菌中的可溶性表达

Soluble Expression of Brucella Outer Membrane Protein Bp26 in E.coli

为了获得布鲁菌外膜蛋白Bp26活性重组蛋白,以布鲁菌M5株菌体DNA为模板,应用PCR方法扩增得到Bp26基因,将该基因片段克隆到载体pET28a中,转化到E.coli BL21(DE3)中,在不同条件下诱导表达,对获得的菌体细胞超声破碎,通过SDS-PAGE鉴定是否为可溶性表达。结果表明,pET28aBp26重组质粒构建成功;终浓度为1.0mmol/L的IPTG在37℃下诱导,重组蛋白表达量最大;含乳糖成分的ZYM-5052培养基和终浓度为1.0mmol/L的IPTG在相同条件下诱导表达蛋白,ZYM-5052培养基可以更好的获得可溶性表达蛋白,为进一步利用大肠埃希菌系统表达生产高活性重组Bp26蛋白提供了一定的参考依据。

In this experiment, in order to obtain the active recombinant protein Bp26 from Brucella outer membrane, the DNA of M5 Brucella was taken as the template. First of all,obtained the Bp26 gene by am- plified PCR. Then,transformed the recombinant plasmids which was made by cloning the Bp26 gene into the vector pET28a into the E. coli BL21 （DE3）. After induced expression under different conditions, bro- ken the obtained bacterial ceils by sonication. At last, identified the soluble expression by SDS-PAGE. The results showed that, the pET28a-Bp26 recombinant plasmids were constructed successfully; induced ex- pression under the conditions of 1.0 mmol/L of IPTG and at 37 ℃ can obtain the maximum expressing content;ZYM-5052 medium containing lactose can obtain better soluble expression protein compared with IPTG at 1.0 mmol/L under the same conditions, which provide the basis for using E. coli expression sys- tem to produce high active recombinant protein BP26.