Wentao Zheng, Yandong Xia, Xue Wang, Shiqing Gao, Diao Zhou, Vinothkannan Ravichandran, Chanjuan Jiang, Qiang Tu, Yulong Yin, Youming Zhang, Jun Fu, Ruijuan Li, Jia Yin
A lack of generic and effective genetic manipulation methods for Pseudomonas has restricted fundamental research and utilization of this genus for biotechnology applications. Phage-encoded homologous recombination (PEHR) is an efficient tool for bacterial genome engineering. This PEHR system is based on a lambda Red-like operon (BAS) from Pseudomonas aeruginosa phage Ab31 and a Rac bacteriophage RecET-like operon (Rec-TEPsy ) from P. syringae pv. syringae B728a and also contains exogenous elements, including the RecBCD inhibitor (Redγ or Pluγ) or single-stranded DNA-binding protein (SSB), that were added to enhance the PEHR recombineering efficiency...
August 25, 2023: Nature Protocols