XY-85 was cultured in marine broth 2,216 (MB; BD Difo™) at 28°C. E. coli strains DH5α and BL21 (DE3) were cultured on Luria–Bertani (LB) agar at 37°C and used as hosts for expressing the protein whose encoding gene was cloned into pET-28a (Ke Lei Biotechnology Co., Ltd). The AHL biosensors Chromobacterium violaceum CV026 and VIR24 (McClean et al., 1997 (link); Someya et al., 2009 (link)), used to detect short-chain (C4 to C8) and long-chain (C8 to C14) AHLs, were maintained on LB agar at 28°C. Pectobacterium caratovorum subsp. caratovorum (Pcc, provided by Dr. Junna He at China Agricultural University) was cultured on Luria–Bertani (LB) agar at 28°C. Kanamycin was added at 50 μg mL−1. C4-HSL, C6-HSL, 3-oxo-C6-HSL, and C8-HSL were purchased from Cayman Chemical Company (Ann Arbor, MI, United States); 3-oxo-C8-HSL, C10-HSL, 3-oxo-C10-HSL, C12-HSL, 3-oxo-C12-HSL, C14-HSL, and 3-oxo-C14-HSL were purchased from Sigma-Aldrich (St. Louis, MO, United States). All of the AHL stock solutions (10 to 500 mM) were prepared in dimethyl sulfoxide (DMSO) and stored under −20°C.
3 oxo c10 hsl
Manufactured by Merck Group
Sourced in United States
3-oxo-C10-HSL is a chemical compound used in laboratory settings. It is a type of quorum sensing molecule that plays a role in bacterial cell-to-cell communication. The core function of 3-oxo-C10-HSL is to act as a signaling molecule, enabling coordination of gene expression and behavior within bacterial populations.
Automatically generated - may contain errors
Lab products found in correlation
C10 hsl, by Merck Group (3 mentions)
3 oxo c8 hsl, by Merck Group (2 mentions)
C6 hsl, by Cayman Chemical (2 mentions)
C12 hsl, by Merck Group (2 mentions)
C14 hsl, by Merck Group (2 mentions)
3 oxo c14 hsl, by Merck Group (2 mentions)
C4 hsl, by Cayman Chemical (2 mentions)
3 oxo c12 hsl, by Merck Group (2 mentions)
3 oxo c6 hsl, by Cayman Chemical (1 mentions)
C8 hsl, by Cayman Chemical (1 mentions)
N decanoyl homoserine lactone, by Merck Group (1 mentions)
N 3 oxo dodecanoyl l homoserine lactone, by Merck Group (1 mentions)
N 3 oxo hexanoyl l homoserine lactone, by Merck Group (1 mentions)
3 oxo c6 hsl, by Merck Group (1 mentions)
Kanamycin, by Takara Bio (1 mentions)
Ampicillin, by Takara Bio (1 mentions)
C4 hsl, by Merck Group (1 mentions)
C6 hsl, by Merck Group (1 mentions)
5 bromo 4 chloro 3 indolyl β d galactoside x gal, by Takara Bio (1 mentions)
Isopropyl β d thiogalactoside iptg, by Takara Bio (1 mentions)
3 protocols using 3 oxo c10 hsl
1
Cultivation and Characterization of AHL Biosensors
2
Degradation of Quorum Sensing Signals by Bacillus spp.
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AHLs degradation activity of cell-free lysates of Bacillus spp. isolates 30b, 32C, and 6 was assessed against different AHL signals using the well diffusion assay. AHL signals tested were; 100 µM C4-HSL (Cayman), 50 µM C6-HSL, 50 µM N- decanoyl-homoserine lactone (C10-HSL), 25 µM N-(3-oxohexanoyl)-L-homoserine lactone (3-oxo-C6-HSL), 25 µM N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C10-HSL), 25 µM 3-oxo-C12-HSL (Sigma-Aldrich). The relative degradation activity was calculated for each AHL signal by calculating the residual AHL from a calibration curve constructed using different concentrations of standard AHL and their induced zone diameter using two AHLs reporter strains; Chromobacterium violaceum (C. violaceum) CV026 and A. tumafaciens KYC55, which respond to short chain and medium-to-long chain AHLs, respectively.39 (link)–42 The maximum degrading activity is defined as 100% relative degradation activity.43 (link)
3
Recombinant Protein Expression and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The TIANprep Midi Plasmid Kit was used for plasmid or genomic DNA extractions and the E com BL21 (DE3) Chemically Competent Cell was the host-expressing cell; the Uniclone One Step Seamless Cloning Kit recombinant plasmid construction kit and the total RNA Extraction Kit were purchased from Genesand company. T4 DNA Ligase, BamH I restriction endonuclease, and other reagents were used to construct the expression vectors. Kanamycin (30 mg/mL), ampicillin (10 mg/mL), 5-Bromo-4-chloro-3-indolyl β-Dgalactoside (X-Gal), and Isopropyl β-D-Thiogalactoside (IPTG) solutions were purchased from TaKaRa and used as the recombinant strain selection medium. The Mag-Beads His-Tag protein puri cation beads were used for the puri cation of the His-Tag target protein, and the target protein concentration was determined by BCA Protein Assay Kit. Western blot analysis was performed on the target protein (PvdQ enzyme). Anti-6 His-Tag mouse monoclonal antibody was used as the primary antibody, HRP-conjugated rabbit anti-mouse IgG was the second antibody, and the EasyBlot ECL kit was used to observe the color reaction. The AHL standard reagents were purchased from Sigma (C4-HSL, C6-HSL, C8-HSL, 3-HSL, 3oxo-C8-HSL, C10-HSL, 3-oxo-C10-HSL, C12-HSL, C14-HSL, 3-oxo-C14-HSL) and dissolved with methanol (HPLC grade, 99.9%), sealed, and stored at -20°C.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!