A Far-Red Emitting Fluorescent Chemogenetic Reporter for In Vivo Molecular Imaging

Far-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially non-fluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae, and chicken embryos. Beyond enabling the genetic encoding of far-red fluorescence, frFAST allowed the design of a far-red chemogenetic reporter of protein–protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.     

A DNA Switch for Detecting Single Nucleotide Polymorphism within a Long DNA Sequence Under Denaturing Conditions

DNA detection is usually conducted under nondenaturing conditions to favor the formation of Watson–Crick base-paring interactions. However, although such a setting is excellent for distinguishing a single-nucleotide polymorphism (SNP) within short DNA sequences (15–25 nucleotides), it does not offer a good solution to SNP detection within much longer sequences. Here we report on a new detection method capable of detecting SNP in a DNA sequence containing 35–90 nucleotides. This is achieved through incorporating into the recognition DNA sequence a previously discovered DNA molecule that forms a stable G-quadruplex in the presence of 7 molar urea, a known condition for denaturing DNA structures. The systems are configured to produce both colorimetric and fluorescent signals upon target binding.     

Genetically Encoded Bioluminescent Indicator for ERK2 Dimer in Living Cells

In this study, a genetically encoded bioluminescent indicator for ERK2 dimer was developed with the split Renilla luciferase complementation method, in which the formation of ERK2 dimer induces a spontaneous emission of bioluminescence in living cells. In response to extracellular stimuli, such as epidermal growth factor (EGF) or 17β‐estradiol (E2), extracellular signal‐regulated kinase 2 (ERK2) is phosphorylated by its upstream kinase MEK, and also phosphorylates its substrates in various regions of the cell, including the nucleus. Phosphorylated ERK2 is led to form its dimer, thereby transporting itself into the nucleus. We demonstrated with the indicator that stimulation with EGF or E2 induces the formation of ERK2 dimer in living MCF‐7 cells. The dynamics of this dimer formation was examined and discussed.     

Application of enzyme-field effect transistor sensor arrays as detectors in a flow-injection analysis system for simultaneous monitoring of medium components. Part II. Monitoring of cultivation processes

Glucose, maltose, sucrose, lactose, ethanol and urea concentrations were monitored simultaneously during the cultivation of Escherichia coli and Saccharomyces cerevisiae by means of enzyme field effect transistors (EnFETs) applying glucose dehydrogenase (GDH), maltase (MAL)/GDH, invertase (INV)/GDH, β-galactosidase (β-GAL)/galactosedehydrogenase (GALDH), alcoholdehydrogenase (ADH)/aldehydedehydrogenase (ALDH), and urease. These enzymes were (co)immobilized on the pH sensitive gates of an eight-FET array. The FET array was integrated in a commercial FIA system.     

酶催化动力学光度法测定锌

近年来酶法在分析测定方面的研究非常活跃，酶法测定锌已有报道，基本为终点测定法。本文中的研究是基于LDH催化下列反应：丙酮酸＋NADH=乳酸＋NAD^＋（氧化型烟酰胺腺嘌呤二核苷酸），锌离子的存在对酶催化反应具有抑制作用，反应速度的变化率与锌离子的浓度呈相关关系，从而可测定试样中的锌离子含量。本方法不需要底物转变完全，与终点测定法相比，可减少测定时间并节省酶量。     

Intramolecular dimers: a new design strategy for fluorescence-quenched probes

Fluorogenic probes dual-labeled with reporter and quencher dyes use a change in fluorescence to monitor biochemical events (e.g., substrate binding or enzyme digestion). Such events change the reporter-quencher distance, which affects fluorescence. Recently, it is has been shown that static quenching through intramolecular dimers is an important mechanism that can sometimes be more efficient than F?rster resonance energy transfer (FRET).     

示差脉冲伏安法测定乳酸脱氢酶活性及纳米微粒生物毒性检测新方法研究

采用示差脉冲伏安法，在乳酸脱氢酶(LDH)酶促体系“丙酮酸盐 + NADH +H⁺ (?) 乳酸盐 + NAD⁺”中，通过检测NAD⁺还原峰电流的变化，测定了不同条件下(不同酶用量、缓冲液pH值以及温度)LDH的活性、酶促体系的米氏常数K_m^NADH以及最大反应速率v_max。并且在最佳实验条件下，通过检测LDH活性的改变，实验考察了3种纳米物质(ZnS，TiO₂(R)和TiO₂(A))对乳酸脱氢酶酶促体系的影响。     

Quinoprotein glucose dehydrogenase modified carbon paste electrode for the detection of phenolic compounds

The development of an amperometric biosensor for the determination of phenolic compounds is described, using quinoprotein glucose dehydrogenase. The enzyme is integrated into carbon paste and its ability to donate electrons to oxidized phenolic compounds during glucose oxidation is exploited. The sensor response is based on electrochemical oxidation of the phenolic compound followed by its enzymatic regeneration when the bulk solution contains glucose and the electrode is potentiostated at +500 mV (vs. Ag/AgCl/0.1 mol/L KCl). As the result of the catalytic analyte regeneration the electrodes offer very sensitive measurements of redox species like p-aminophenol and hydroquinone and catecholamines such as epinephrine, norepinephrine, and dopamine. The sensor performance is characterized for the different substrates. Highest sensitivity is achieved for p-aminophenol which could be determined at sub-nanomolar level.     

薄层扫描法测定冬眠过程中刺猬活性组织酶的周期变化

薄层扫描法测定冬眠过程中刺猬活性组织酶的周期变化冯金城（天津师范大学天津３０００７４）孙金生（天津水产研究所天津３００２２１）１前言为了跟踪观察刺猬在冬眠过程中体内乳酸脱氢酶的周期性变化，利用薄层扫描仪测定了其相关的组织同工酶的差异。实践证明，此法简...