抗菌肽Dermadistinctin-Q1的原核表达及其生物活性鉴定

摘要

This experiment expressed antibacterial peptide Dermadistinctin-Q1 (DDQ1)in prokaryotic system and de-tected its biological activity in vitro,the splice of genes by overlap extension was applied,and then the complete gene se-quence of DDQ1 was obtained by multistep PCR.The engineering bacteria was constructed by prokaryotic expression vector pET-32a (+).The optimization of fusion protein expression conditions came ture via optimizing the inducible expression time and the final concentration of IPTG.Antibacterial peptide DDQ1 was separated from the fusion protein by enterokinase and was purified by Ni ion affinity chromatography.The further detection of antibacterial peptide DDQ1 biological activitiy was performanced.The results showed that antibacterial peptide DDQ1 had broad-spectrum antibacterial activity,as well as a certain thermal-tolerance and acid-tolerance.Therefore,it was a kind of extremely potential new drug.%试验对抗菌肽Dermadistinctin-Q1（DDQ1）进行原核表达，并对其生物活性进行检测。采用重叠区扩增基因拼接法，多步PCR获得抗菌肽DDQ1全基因，并使用原核表达载体pET-32a（+）构建工程菌。通过优化诱导表达时间和异丙基硫代半乳糖苷（IPTG）终浓度，实现融合蛋白表达条件最佳化。利用肠激酶从融合蛋白中分离抗菌肽DDQ1后，使用Ni离子亲和层析法进行纯化。进一步的生物活性检测表明，抗菌肽DDQ1具有广谱抗菌活性和一定的热、酸耐受性，是一类极具开发潜力的新型药物。