Modulation of OMV Production by the Lysis Module of the DLP12 Defective Prophage of Escherichia coli K12
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and General Growth Conditions
2.2. Plasmid Construction
2.3. General Procedures
2.4. Biosensor Assay
2.5. OMV Purification
2.6. SDS-PAGE and Immunoblot Analysis
2.7. DiO Staining
2.8. Statistical Analyses
E. coli Strain | Relevant Characteristics | Source/Reference |
---|---|---|
DH10b | E. coli K12 | [40] |
MG1655 | E. coli K12, F-, λ-, ilvG-, rfb-50, rph-1 | [41] |
ULS153 | ΔlacZYA derivative of MG1655 | [30] |
ULS153 ΔDlm | ULS153 mutant defective in DLP12 operon | This study |
ULS153 ΔdegP | ULS153 mutant defective in degP gene | This study |
ULS153 ΔS | ULS153 mutant defective in essD gene | This study |
ULS153 ΔR | ULS153 mutant defective in ybcS gene | This study |
ULS153 ΔRz | ULS153 mutant defective in rzpD/rzoD genes | This study |
ULS153 ΔSR | ULS153 mutant defective in essD and ybcS genes | This study |
ULS153 ΔRRz | ULS153 mutant defective in ybcS and rzpD/rzoD genes | This study |
ULS153 ΔSRz | ULS153 mutant defective in essD and rzpD/rzoD genes | This study |
Plasmid Name | Relevant Characteristics | Source/Reference |
pACYC184 | Low/Medium copy number cloning vector | [40] |
pAC2.10 | pACYC184 derivate carrying the DLP12 operon | This study |
pKD46 | Red recombinase expression plasmid | [31] |
pKD3 | Template plasmid carrying a CmR gene with FLP recognition target sequence | [31] |
pCP20 | Temperature sensitive replicon carrying the yeast Flp recombinase gene | [31] |
pAG4 | High copy number plasmid carrying the lux operon | [33] |
pAGPDlm | pAG4 derivate carrying the DLP12 promoter region of MG1655 strain upstream lux operon | This study |
pAGPlac | pAG4 derivate carrying the lac promoter region of MG1655 strain upstream lux operon | This study |
3. Results
3.1. The SRRz/Rz1 Operon Regulates OMV Production
3.2. Defining the Role of the Single Genes of the SRRz/Rz1 Operon
3.3. Stress Factors Modulate the Expression of SRRz/Rz1 Operon
4. Discussion
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Pasqua, M.; Zennaro, A.; Trirocco, R.; Fanelli, G.; Micheli, G.; Grossi, M.; Colonna, B.; Prosseda, G. Modulation of OMV Production by the Lysis Module of the DLP12 Defective Prophage of Escherichia coli K12. Microorganisms 2021, 9, 369. https://doi.org/10.3390/microorganisms9020369
Pasqua M, Zennaro A, Trirocco R, Fanelli G, Micheli G, Grossi M, Colonna B, Prosseda G. Modulation of OMV Production by the Lysis Module of the DLP12 Defective Prophage of Escherichia coli K12. Microorganisms. 2021; 9(2):369. https://doi.org/10.3390/microorganisms9020369
Chicago/Turabian StylePasqua, Martina, Alessandro Zennaro, Rita Trirocco, Giulia Fanelli, Gioacchino Micheli, Milena Grossi, Bianca Colonna, and Gianni Prosseda. 2021. "Modulation of OMV Production by the Lysis Module of the DLP12 Defective Prophage of Escherichia coli K12" Microorganisms 9, no. 2: 369. https://doi.org/10.3390/microorganisms9020369