Fernandes de Brito, Luciana; Schultenkämper, Kerstin; Passaglia, Luciane; Wendisch, Volker F.: CRISPR interference-based gene repression in the plant growth promoter Paenibacillus sonchi genomovar Riograndensis SBR5. In: Applied Microbiology and Biotechnology. Jg.104. 2020, S. 5095-5106
Inhalt
- Strains, plasmids, and oligonucleotides
- Media and cultivation conditions
- Plasmid construction and preparation of recombinant strains
- Quantification of sporulation
- Colony phenotyping of recombinant P.sonchi strains
- Quantification of biofilms
- GFPUV fluorescence measurement by flow cytometry
- RNA isolation and qRT-PCR
- Determination of YdjJ specific activity
- Results
- CRISPRi-mediated repression of plasmid-borne GFPUV
- Effect of ydjJ gene repression on P.sonchi SBR5 grown in d-sorbitol
- CRISPRi targeting sporulation regulatory gene spo0A decreased its mRNA level
- Hemocytometer spore counts of P.sonchi cells targeting spo0A and yaaT
- Colony phenotyping analysis
- Effect of CRISPRi on biofilm formation by P.sonchi
- Discussion
- References