蜂毒肽与死亡素杂合肽(MT)在大肠杆菌中融合表达的研究

Fusion Expressing of Melitin and Thanantin Heteropeptide(MT) in E.coli

人工合成的蜂毒肽与死亡素的杂合肽DNA序列,经聚合酶链式反应(PCR)扩增后得到带有BamHI和XhoI限制性酶切位点的序列.将此基因克隆至载体pET-32a,成功构建了重组质粒pET32a-MT.测序验证后转化大肠杆菌BL21(DE3),37℃IPTG诱导,获得高效表达,融合蛋白经Ni柱亲和纯化后用肠激酶切下杂合肽,杯碟法检验酶切产物具有抑菌活性。

The eDNA sequences of melitin and thanatin heteropeptide was synthesized by PCR, then cloned into the fusion vector pET-32a and transformed into E. coli BL21 （DE3）. Furthermore the expected sequences were checked. Induced by IPTG at 37 ℃, the fusion protein was expressed. The protein was purified by HiTrapTM chelating Kit and then digested by rE kit , then the inhibition activity of the MT peptide was tested by disc methods, and which exhibit anetimierobial activity.