基础医学与临床 ›› 2010, Vol. 30 ›› Issue (10): 1081-1084.

• 研究论文 • 上一篇    下一篇

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

王浩 周建 罗理 董碧蓉   

  1. 解放军第452医院 四川大学华西医院
  • 收稿日期:2009-09-14 修回日期:2010-02-24 出版日期:2010-10-05 发布日期:2010-10-05
  • 通讯作者: 董碧蓉

Human α-defensin-3 enhanced Ciprofloxacin antibiotic activity to resistant strains of Pseudomonas aeruginosa

Hao WANG, Jian ZHOU, Li LUO, Bi-rong DONG   

  1. The People's Liberation Army No.452 Hospital West China Hosipital, Sichuan university
  • Received:2009-09-14 Revised:2010-02-24 Online:2010-10-05 Published:2010-10-05
  • Contact: Bi-rong DONG

摘要: 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,并PCR扩增目的基因,将其克隆入pET32a(+)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MIC值和FIC值检测重组防御素蛋白生物活性。结果 成功构建出pET32a(+)/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都小于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。

关键词: 防御素, 基因重组, 铜绿假单胞菌, FIC

Abstract: Objective we got a lot of recombinant human α-defensin-3 by genetic engineering method, and studied whether to increase ciprofloxacin antibiotic activity to resistant strains of Pseudomonas aeruginosa. Method RT-PCR retroviral human blood leukocytes of the total RNA, and PCR amplification of the target gene, cloned into pET32a (+) plasmid. Transformed into the recombinant E. coli and the affinity chromatography purified protein. By calculating HNP-3 and ciprofloxacin MIC value and FIC detection of recombinant protein biological activity. Results Construction of a pET32a (+) / HNP-3 original expression vector, and the expression of protein was determined the purpose of protein. The FIC value of combination of HNP-3 and ciprofloxacin is less than 1.0. Conclusion human α-defensin-3 enhanced ciprofloxacin antibiotic activity to resistant strains of Pseudomonas aeruginosa.

Key words: Human neutrophil peptide, Gene recombination, Pseudomonas aeruginosa., FIC