Mrr from Escherichia coli K12 is a type IV restriction endonuclease whose role is to recognize and cleave foreign methylated DNA. Beyond this protective role, Mrr can inflict chromosomal DNA damage that elicits the SOS response in the host cell upon heterologous expression of specific methyltransferases such as M.HhaII, or after exposure to high pressure (HP). Activation of Mrr in response to these perturbations involves an oligomeric switch that dissociates inactive homo-tetramers into active dimers. Here we used scanning number and brightness (sN&B) analysis to determine in vivo the stoichiometry of a constitutively active Mrr mutant predicted to be dimeric and examine other GFP-Mrr mutants compromised in their response to either M.HhaII activity or HP shock. https://www.selleckchem.com/products/ABT-263.html We also observed in vitro the direct pressure-induced tetramer dissociation by HP fluorescence correlation spectroscopy of purified GFP-Mrr. To shed light on the linkages between subunit interactions and activity of Mrr and its variants, we built a structural model of the full-length tetramer bound to DNA. Similar to functionally related endonucleases, the conserved DNA cleavage domain would be sequestered by the DNA recognition domain in the Mrr inactive tetramer, dissociating into an enzymatically active dimer upon interaction with multiple DNA sites.Olive oil is more preferred than other vegetable oils because of the increasing health concern among people throughout the world. The major hindrance in large-scale production of olive oil is olive fruit pests which cause serious economic damage to the olive orchards. This requires careful monitoring and timely application of suitable remedies before pest infestation. Herein we demonstrate efficacious utilization of covalently functionalized β-cyclodextrinylated MEMS devices for selective and sensitive detection of female sex pheromone of olive fruit pest, Bactocera oleae. Two of the MEMS devices, silicon dioxide surface-micromachined cantilever arrays and zinc oxide surface-microfabricated interdigitated circuits, have been used to selectively capture the major pheromone component, 1,7-dioxaspiro[5,5]undecane. The non-covalent capture of olive pheromones inside the β-cyclodextrin cavity leads to the reduction of resonant frequency of the cantilevers, whereas an increase in resistance has been found in case of zinc oxide derived MEMS devices. Sensitivity of the MEMS devices towards the olive pheromone was found to be directly correlated with the increasing availability of β-cyclodextrin moieties over the surface of the devices and thus the detection limit of the devices has been achieved to a value as low as 0.297 ppq of the olive pheromone when the devices were functionalized with one of the standardized protocols. Overall, the reversible usability and potential capability of the suitably functionalized MEMS devices to selectively detect the presence of female sex pheromone of olive fruit fly before the onset of pest infestation in an orchard makes the technology quite attractive for viable commercial application.Traditional approaches for nucleic acids detection require prior amplification of target genes, while nanomaterials-aided DNA biosensors are very magnificent but still suffer from the nanomaterial acquirement and limited sensitivity (above picomolar level). Herein, fullerenol C60(OH)25, a representative fullerene derivative, was employed as a photoelectrochemical (PEC) nanoprobe to achieve discrimination and ultrasensitive detection of amplification-free single-stranded DNA (ssDNA) down to sub-femtomolar level. The bonded hydroxyl groups with intense density endowed fullerenol to directly recognize and capture ssDNA-AuNPs via the hydrogen bonding interactions (H-bonds), leading to a sharply decreased photocurrent with quenching efficiency up to 85%, which could be attributed to the photo-generated electrons on the conduction band of fullerenol (-4.66 eV) preferentially migrating to the Fermi level of AuNPs (-5.1 eV) rather than the electrode. In the presence of target gene (mutant human p53 gene fragment), the H-bonds between fullerenol and ssDNA were competitively depleted during the base pairing process of complete hybridization between ssDNA and target, making double-stranded DNA-AuNPs (dsDNA-AuNPs) depart so that the photocurrent powerfully recovered. On basis of the photocurrent variation before and after target introduction, this proposed simple, rapid and ultrasensitive PEC biosensor for amplification-free target gene detection illustrated a wide liner ranged from 1 fM to 100 pM and a detection limit of 0.338 fM. This work presented an ingenious strategy for the discrimination and ultrasensitive detection of nucleic acids, and the well-designed PEC biosensor was further conducive to the impetus of clinic diagnostics.Bacterial infection is a growing public health concern and causes a huge medical and financial burden. It is of significance to efficiently construct multifunctional platforms for bacterial point-of-care testing (POCT) and elimination. Herein, near-infrared (NIR) light-responded vancomycin-doped prussian blue nanoparticles (PB-VANNPs) with high efficient photothermal conversion was synthesized for binding, dual-mode portable detection, and elimination of bacteria. The PB-VANNPs can bind to the surface of Gram-positive bacteria such as Staphylococcus aureus (S. aureus), forming complex of PB-VANNPs/S. aureus. After being centrifugated, the suspension solution of PB-VANNPs can stimulate perfluorohexane (PFH) to rapidly release oxygen (O2) under NIR irradiation. Thus, the bacteria can be sensitively detected with portable pressure meter as signal reader, reporting a limit of detection (LOD) of 1.0 CFU mL-1. On the other side, the sediment of PB-VANNPs/S. aureus can be detected via thermal camera, reporting a LOD of 1.0 CFU mL-1. Interestingly, the bacteria can be effectively inactivated with the local temperature elevation during temperature-based detection. The antibacterial efficiency reaches as high as 99.8%. The developed multifunctional nanoplatform not only provides a straightforward "mix-then-test" way for portable detection of bacteria with high sensitivity, also realizes high efficiency elimination of bacteria simultaneously. The developed strategy was further applied for promoting wound healing of bacteria-infected mice.