【摘要】 为了在大肠杆菌中表达杂合抗菌肽Magainin-Thanatin(MT).通过大肠杆菌密码子偏爱性优化两抗菌肽基因序列,SOE-PCR技术构建克隆载体并测序;将其转化至表达菌株Escherichia coli BL21(DE3)中构建基因工程菌,表达产物做抑菌活性分析.结果在本研究中经SOE-PCR拼接后得到了杂合肽片段,成功构建了表达载体pGEX-6P-1-MT;转化后得到了重组菌株pGEX-6P-1-MT/BL21(DE3);获得诱导产物分子质量约3.35kDa;其对大肠杆菌DH5α、金黄色葡萄球菌等均有抑制作用.这表明杂合肽MT可以在大肠杆菌BL21(DE3)中融合分泌表达且具有抗菌活性.更多还原

【Abstract】 In order to express the hybrid antimicrobial peptide Magainin-Thanatin(MT)in Escherichia coli.The antimicrobial peptide gene sequence was optimized by the codon preference of Escherichia coli.SOE-PCR was used to construct the cloning vector.The vector was converted to the expressed strain Escherichia coli BL21(DE3)to construct a genetically engineered strain.The expression product was verified for antimicrobial activity.In this study,heterozygous peptide fragments were obtained by SOE-PCR splicing,and the expression vector pGEX-6 P-1-MT was successfully constructed.The recombinant strain pGEX-6 P-1-MT/BL21(DE3)was achieved after transformation.The molecular weight of the induced product was about 3.35 kDa.The target protein has an inhibitory effect on Escherichia coli DH5αand Staphylococcus aureus.This indicates that the hybrid peptide MT can be secreted and expressed in Escherichia coli BL21(DE3)and has antibacterial activity.更多还原