A recombineering system for Bacillus subtilis based on the native phage recombinase pair YqaJ/YqaK

Bacillus subtilis as a host for natural product discovery and engineering of biosynthetic gene clusters

“…A recently developed recombineering system for B. subtilis , based on the recombinase pair YqaJ/YqaK from a B. subtilis 168 prophage, can be used to insert large DNA sequences as well as introduce gene deletions in BGCs. 273 To achieve this, the transformation efficiency of B. subtilis was enhanced through overexpression of the ComK regulator. The system relies on the co-expression of yqaJK and comK , and utilizes a double-stranded DNA substrate with 100 base pair homology arms and a phosphorothioate modification at the 5′-end of the lagging targeting strand to improve recombination efficiency.…”

“…274 Taken together, the YqaJK system proved to be superior to other recombinase systems previously used in B. subtilis , making it an efficient genome manipulation tool for this species. 273…”