Red重组系统在克雷伯氏肺炎杆菌中的应用研究

Application of Red Recombination System in Klebsiella Pneumoniae

通过改造pKD46与pCP20质粒为含四环素抗性标记的pKD46-Tc与pCP20-Tc重组质粒,初步建立了一套能在K.pneumoniae菌株中运用的Red重组系统。以荚膜基因中的高保守基因wzi为敲除对象,导入同源臂为250 bp的打靶片段△wzi::FK,转化后得到6个重组子,初步确定敲除了K.pneumoniae菌株的荚膜基因wzi。

Through constructing the plasmid pKD46-Tc and pCP20-Tc which contained tetracycline resistance marker,a Red recombination system in Klebsiella pneurnoniae was established. Using the gene zvzi, a highly conserved homologous gene of capsule gene, as a target gene, a fragment called Awzi： ：FK which contained 250 bp homologous segments was constructed and it was transformed into Klebsiella pneumoniae/ pKD46-Tc, then resulted six recombinants. The capsule gene wzi was preliminarily verified having been knockout by using the Red recombination system in Klebsiella pneumoniae.