Extremely low levels of "leaky" expression of genes in bacterial protein expression systems can severely curtail cell viability when expressed proteins are toxic. A general method for sensitive detection of such expression is lacking. Here, we present a method based on microscopic visualization of a fluorescent "reporter" protein (RFP-HU-A) constructed by fusing red fluorescent protein (RFP) to the N-terminus of a nucleoid-associated, histone-like DNA-binding protein, HU-A. Localization of RFP-HU-A within nucleoids facilitates detection, quantitation, and characterization of leaky expression at the single-cell level.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.ab.2015.06.011 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Comparing sputum, nasopharyngeal swabs, and combined samples for respiratory bacterial detection using multiplex PCR.
Microbiol Spectr
January 2025
Department of Laboratory Medicine, Korea University Anam Hospital, Korea University College of Medicine, Seoul, South Korea.
Unlabelled: Respiratory tract infections are major global health issues that require rapid and accurate diagnostic methods. Multiplex quantitative PCR (qPCR) is commonly used for pathogen detection in respiratory samples. However, the optimal specimen selection for detecting bacterial pathogens is not well-explored.
View Article and Find Full Text PDFSynaptic enrichment and dynamic regulation of the two opposing dopamine receptors within the same neurons.
Elife
January 2025
Graduate School of Life Sciences, Tohoku University, Sendai, Japan.
Dopamine can play opposing physiological roles depending on the receptor subtype. In the fruit fly , and encode the D- and D-like receptors, respectively, and are reported to oppositely regulate intracellular cAMP levels. Here, we profiled the expression and subcellular localization of endogenous Dop1R1 and Dop2R in specific cell types in the mushroom body circuit.
View Article and Find Full Text PDFSimultaneous measurement of fentanyl, fentanyl analogues and other drugs of abuse by multiplex bead assay.
Toxicol Mech Methods
January 2025
Centers for Disease Control and Prevention, Division of Science Integration, Risk Evaluation Branch, National Institute for Occupational Safety and Health, Cincinnati, OH, USA.
Quantification of illicit drugs and controlled substances, in urine or as surface contamination, is often performed using expensive analytical techniques such as liquid chromatography with tandem mass spectrometry (LC-MS/MS). A time and cost-effective semi-quantitative surface-wipe and urine screening multiplex immunoassay for fentanyl and its analogues was developed in this investigation. We previously created a surface wipe multiplex immunoassay for methamphetamine, caffeine, cocaine, tetrahy-drocannabinol (THC) and oxycodone.
View Article and Find Full Text PDFInvestigating the role of IL-6 in the pathogenesis of systemic lupus erythematosus: Insights from bone marrow mesenchymal stem cells.
Lupus
January 2025
Department of Dermatology, Nanfang Hospital, Southern Medical University, Guangzhou, People's Republic of China.
Background: Systemic lupus erythematosus is a common autoimmune disease. Studies have suggested that defective stem cells could be involved in the pathogenesis of systemic lupus erythematosus, which leads to changes in the function of immune cells. By observing the cell morphology, autophagy, and senescence of bone marrow mesenchymal stem cells (BMSCs) from lupus mice and normal controls, this study investigated the role of IL-6 in autophagy and senescence of BMSCs and explored relevant mechanisms.
View Article and Find Full Text PDFExogenous Coreactant-Free Electrocatalytic Reactive Oxygen Species-Driven Dual-Signal Molecularly Imprinted Electrochemiluminescence Sensor for the Detection of Trenbolone.
Anal Chem
January 2025
The Education Ministry Key Lab of Resource Chemistry, Joint International Research Laboratory of Resource Chemistry, Ministry of Education, Shanghai Frontiers Science Center of Biomimetic Catalysis, Shanghai Key Laboratory of Rare Earth Functional Materials, College of Chemistry and Materials Science, Shanghai Normal University, Shanghai 200234, China.
Conventional dual-signal electrochemiluminescence (ECL) sensors feature high sensitivity and reliability, but the involvement of coreactants inevitably results in a complex configuration and shows reproducibility risk. Here, we propose an exogenous coreactant-free dual-signal platform, comprising luminol (anodic luminophore), CdSe quantum dots (cathodic luminophore), and CoO/TiC electrocatalyst (coreaction promoter). At different redox potentials, CoO/TiC induces water oxidation and oxygen reduction to produce OH and O radicals, which subsequently drive cathodic and anodic ECL emission, respectively.
View Article and Find Full Text PDFWant 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!