Recombineering-Mediated Genome Editing in Burkholderiales Strains.

Methods Mol Biol

Helmholtz International Lab for Anti-infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao, People's Republic of China.

Published: May 2022

Red/ET recombineering is primarily mediated by the E. coli recombinase pair Redα/Redβ from λ phage or RecE/RecT from Rac prophage, which is applied in E. coli and also closely related Gram-negative bacteria for efficient genome editing. However, some distant bacterial species like Burkholderiales strains require host-specific Redα/Redβ recombinase pair for highly efficient genome editing. A pair of recombinases Redαβ7029 from the Burkholderiales strain DSM 7029, recently identified as Schlegelella brevitalea, were identified for efficient genetic manipulation in the native strain and several other Burkholderiales strains. In this chapter, we describe a detailed protocol for genome engineering in Burkholderiales strains via the Redγ-Redαβ7029 recombineering and Cre/loxP site-specific recombination.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-2233-9_3DOI Listing

Publication Analysis

Top Keywords

burkholderiales strains
16
genome editing
12
recombinase pair
8
efficient genome
8
burkholderiales
5
recombineering-mediated genome
4
editing burkholderiales
4
strains
4
strains red/et
4
red/et recombineering
4

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!