人防御素3增强环丙沙星对铜绿假单胞菌耐药株杀菌活性

Human α-defensin-3 enhances ciprofloxacin antibiotic activity to resistant strains of Pseudomonas aeruginosa

目的使用基因工程的方法获得大量重组人防御素3(HNP-3),并研究其是否能增强环丙沙星对铜绿假单胞菌耐药株杀菌活性。方法RT-PCR反转录人血白细胞的总RNA,PCR扩增目的基因,将其克隆入pET32 a(+)质粒中。转化重组质粒入大肠杆菌,表达大量蛋白,经亲和层析纯化目的蛋白。通过计算HNP-3与环丙沙星的M IC值和FIC值来检测重组防御素蛋白生物活性。结果成功构建出pET32 a(+)/HNP-3原核表达载体,证明表达的蛋白为目的蛋白。HNP-3和环丙沙星联合后的FIC均小于1.0。结论人防御素3增强抗生素对铜绿假单胞菌耐药株杀菌活性。

Objective To explore potential enhancement of ciprofloxcin to kill resistant strains of Pseudomonas aeruginosa.Methods RT-PCR retroviral human blood leukocytes of the total RNA,and PCR amplification of the target gene were performed and cloned into pET32a（＋） plasmid and then transformed into the recombinant E.coli and obtained the affinity chromatography purified protein,by calculating HNP-3 and ciprofloxacin MIC value and FIC to detect the recombinant protein biological activity.Results Construction of a pET32a（＋） / HNP-3 original expression vector,and the expression of protein was proved to be purpose protein.The FIC value of combination of HNP-3 and ciprofloxacin was less than 1.0.Conclusion Human α-defensin-3 enhanced ciprofloxacin activity to kill the resistant strains of Pseudomonas aeruginosa.