In vivo expression technology for selection of bacterial genes specifically induced in host tissues.

abstract：:The current treatment of neurological and psychiatric diseases is far beyond being satisfactory. In addition to highly complex disease mechanisms, the blood-brain barrier (BBB) also remains as a challenge by limiting the delivery of the majority of currently available therapeutics to the central nervous system. Severa...

journal_title：Methods in enzymology

authors： Yemişci M,Gürsoy-Özdemir Y,Caban S,Bodur E,Capan Y,Dalkara T

更新日期：2012-01-01 00:00:00

abstract：:Cancer has become the leading cause of death in the developed world and has remained one of the most difficult diseases to treat. One of the difficulties in treating cancer is that conventional chemotherapies often have unacceptable toxicities toward normal cells at the doses required to kill tumor cells. Thus, the de...

journal_title：Methods in enzymology

authors： Cantor JR,Panayiotou V,Agnello G,Georgiou G,Stone EM

更新日期：2012-01-01 00:00:00

abstract：:Because polysaccharides have very complicated chemical structures constructed by a great diversity of monosaccharide residues and glycosidic linkages, enzymatic approaches have been identified as powerful tools to precisely synthesize polysaccharides as the reactions progress in highly controlled regio- and stereoarra...

journal_title：Methods in enzymology

authors： Kadokawa JI

更新日期：2019-01-01 00:00:00

abstract：:The neutral arachidonic acid derivatives N-arachidonoyl ethanolamine (anandamide or AEA), and 2-arachidonoylglycerol (2-AG) have been identified as endogenous ligands for the cannabinoid receptors. Additionally, these compounds have been identified as substrates of the second isoform of the cyclooxygenase enzyme (COX-...

journal_title：Methods in enzymology

authors： Kingsley PJ,Marnett LJ

更新日期：2007-01-01 00:00:00

abstract：:The genome integrity of living organisms is constantly threatened by endogenous cellular metabolic processes and environmental agents. To quantify these low, physiologically relevant levels of DNA damage, a single-cell gel electrophoresis (comet) combined with strand-specific fluorescence in situ hybridization (FISH)-...

journal_title：Methods in enzymology

authors： Mondal M,Guo J

更新日期：2017-01-01 00:00:00

abstract：:The field of therapeutic application of the A₃ adenosine receptor (A₃AR) antagonists represents a rapidly growing and intense area of research in the adenosine field. Even if there are currently no A₃AR antagonists in clinical phases, in light of the plethora of biological effects attributed to A₃ARs, substantial effo...

journal_title：Methods in enzymology

authors： Paoletta S,Federico S,Spalluto G,Moro S

更新日期：2010-01-01 00:00:00

abstract：:Glycoproteins are a major class of glycoconjugates displaying a variety of mutual interactions between glycan and protein moieties that ultimately affect molecular organization. Modulation of the pendant glycan structures is important in tuning the functions of glycoproteins. Here we discuss structural aspects and som...

journal_title：Methods in enzymology

authors： Barb AW,Borgert AJ,Liu M,Barany G,Live D

更新日期：2010-01-01 00:00:00

abstract：:Neurofilament biology is important to understanding structural properties of axons, such as establishment of axonal diameter by radial growth. In order to study the function of neurofilaments, a series of genetically modified mice have been generated. Here, we describe a brief history of genetic modifications used to ...

journal_title：Methods in enzymology

authors： Jones MR,Villalón E,Garcia ML

更新日期：2016-01-01 00:00:00

abstract：:The human cannabinoid receptor, CB1, has been difficult to purify in a functional form, hampering structural and biophysical studies. Here, we present our approaches for obtaining pure, detergent solubilized, functional CB1. We also discuss our site-directed fluorescence labeling (SDFL) methods for identifying differe...

journal_title：Methods in enzymology

authors： Fay JF,Farrens DL

更新日期：2017-01-01 00:00:00

abstract：:Aging that involves contact with dying yeast cells is one of the differential processes between sparkling and still wine production. The release of the products of autolysis during this aging step is fundamental for the quality of sparkling wines made by the traditional method. These cells undergo an autolysis process...

journal_title：Methods in enzymology

authors： Cebollero E,Rejas MT,González R

更新日期：2008-01-01 00:00:00

abstract：:Embryonic stem (ES) cells are cells derived from the inner cell mass of a blastocyst stage embryo. These self-renewing multipotent cells are able to differentiate to the three embryonic germ layers, the endoderm, ectoderm, and mesoderm, and are thus able to produce virtually all cell types. The ES cell capacity to gen...

journal_title：Methods in enzymology

authors： Goldberg-Cohen I,Beck G,Ziskind A,Itskovitz-Eldor J

更新日期：2006-01-01 00:00:00

abstract：:Desmoplakin (DP) is an obligate component of desmosomes, where it links the desmosomal cadherin/plakoglobin/plakophilin assembly to intermediate filaments. DP contains a large amino-terminal domain (DPNT) that binds to the cadherin/plakoglobin/plakophilin complex, a central coiled-coil domain that dimerizes the molecu...

journal_title：Methods in enzymology

authors： Choi HJ,Weis WI

更新日期：2016-01-01 00:00:00

abstract：:A number of different analytical techniques are now available for the isolation of apoA-I, apoA-II, and apoA-IV. The choice of a particular technique is dependent on the instrumentation available, and the quantity of isolated apolipoprotein required. The isolation and characterization of the separate isoforms and the ...

journal_title：Methods in enzymology

authors： Brewer HB Jr,Ronan R,Meng M,Bishop C

更新日期：1986-01-01 00:00:00

abstract：:Reliable modeling of RNA tertiary structures is key to both understanding these structures' roles in complex biological machines and to eventually facilitating their design for molecular computing and robotics. In recent years, a concerted effort to improve computational prediction of RNA structure through the RNA-Puz...

journal_title：Methods in enzymology

authors： Cheng CY,Chou FC,Das R

更新日期：2015-01-01 00:00:00

abstract：:To maintain cellular homeostasis, the levels of transmembrane receptors found on the plasma membrane must be tightly regulated. Endocytosis of activated receptors and the eventual degradation of these transmembrane proteins in the lysosome serve a vital role in maintaining the plasma membrane receptor levels as well a...

journal_title：Methods in enzymology

authors： Davies BA,Carney DS,Horazdovsky BF

更新日期：2005-01-01 00:00:00

abstract：:Chloroplast genomes of land plants and algae contain generally between 100 and 150 genes. These genes are involved in plastid gene expression and photosynthesis and in various other tasks. The function of some chloroplast genes is still unknown and some of them appear to be essential for growth and survival. Repressib...

journal_title：Methods in enzymology

authors： Ramundo S,Rochaix JD

更新日期：2015-01-01 00:00:00

abstract：:The cell membrane isolation procedure we developed here can be scaled up from four to several hundred plates of cultured cells. Transmission electron microscopy, membrane marker enzyme analysis, binding study, EGF-dependent receptor autophosphorylation, and Western blots all demonstrate the biological activity of the ...

journal_title：Methods in enzymology

authors： Lin PH,Selinfreund R,Wharton W

更新日期：1991-01-01 00:00:00

abstract：:As for other GPCRs, the oligomerization of glycoprotein hormone receptors (GPHRs) appears as critical event for receptor function. By means of modern techniques based on the BRET or FRET principle, GPHR oligomerization has been reported to explain several physiological and pathological conditions. In particular, the p...

journal_title：Methods in enzymology

authors： Bonomi M,Persani L

更新日期：2013-01-01 00:00:00

abstract：:Experimental screening for protein-ligand interactions is a central task in drug discovery. Nuclear magnetic resonance (NMR) spectroscopy enables the determination of binding affinities, as well as the measurement of structural and dynamic parameters governing the interaction. With traditional liquid-state NMR relying...

journal_title：Methods in enzymology

authors： Kim Y,Hilty C

更新日期：2019-01-01 00:00:00

abstract：:Critical to the advancement of tumor immunotherapy is the reliable identification of responders and the quantification of the tumor-specific immune response elicited by treatments. In this regard, Enzyme-Linked Immunospot assay (ELISpot) is an ideal monitoring technique due to its high sensitivity, ease of execution a...

journal_title：Methods in enzymology

authors： Negri D,Sestili P,Borghi M,Ciccolella M,Bracci L

更新日期：2020-01-01 00:00:00

abstract：:Lysosomes are specialized organelles for protein recycling and as such are involved in the terminal steps of autophagy. However, it has become evident that lysosomes also play an important role in the progression of apoptosis. This latter function seems to be dependent on lysosomal proteases, which need to be released...

journal_title：Methods in enzymology

authors： Ivanova S,Repnik U,Bojic L,Petelin A,Turk V,Turk B

更新日期：2008-01-01 00:00:00

abstract：:The Ras gene family has been implicated in the development of many human epithelial cancers. Mutations in K-ras or its downstream mediator BRAF have been detected in about two thirds of low-grade serous carcinomas and borderline serous tumors; mutations in K-ras are also often present in benign and invasive mucinous o...

journal_title：Methods in enzymology

authors： Rosen DG,Yang G,Bast RC Jr,Liu J

更新日期：2006-01-01 00:00:00

abstract：:In this chapter, we motivate the need for photoactivatable NO donor molecules and give a brief survey of the existing chemical tools in the field. We then provide detailed protocols for the synthesis and validation of a near-infrared light-activated NO donor molecule, photoNOD-1, developed in our research group. With ...

journal_title：Methods in enzymology

authors： Zhou EY,Knox HJ,Reinhardt CJ,Partipilo G,Chan J

更新日期：2020-01-01 00:00:00

abstract：:The ability to examine gene regulation in living cells has been greatly enabled by the development of chromatin immunoprecipitation (ChIP) methodology. ChIP captures a snapshot of protein-DNA interactions in vivo and has been used to study interactions in bacteria, yeast, and mammalian cell culture. ChIP conditions va...

journal_title：Methods in enzymology

authors： Davis SE,Mooney RA,Kanin EI,Grass J,Landick R,Ansari AZ

更新日期：2011-01-01 00:00:00

abstract：:Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the ...

journal_title：Methods in enzymology

authors： Lyumkis D,Moeller A,Cheng A,Herold A,Hou E,Irving C,Jacovetty EL,Lau PW,Mulder AM,Pulokas J,Quispe JD,Voss NR,Potter CS,Carragher B

更新日期：2010-01-01 00:00:00

abstract：:This chapter presents methodologies for RNA extraction from soils coupled with competitive reverse transcription-polymerase chain reaction and capillary electrophoresis techniques. Combined, these approaches provide new capabilities to quantify gene expression in different environments and can aid our understanding of...

journal_title：Methods in enzymology

authors： Semrau JD,Han JI

更新日期：2005-01-01 00:00:00

abstract：:Genome editing using the Cas9 endonuclease of Streptococcus pyogenes has demonstrated unprecedented efficacy and facility in a wide variety of biological systems. In zebrafish, specifically, studies have shown that Cas9 can be directed to user-defined genomic target sites via synthetic guide RNAs, enabling random or h...

journal_title：Methods in enzymology

authors： Gonzales AP,Yeh JR

更新日期：2014-01-01 00:00:00

abstract：:Hybrid proteins or chimeras are generated by recombination of protein fragments. In the course of evolution, this mechanism has led to major diversification of protein folds and their functionalities. Similarly, protein engineers have taken advantage of this attractive strategy to build new proteins. Methods that use ...

journal_title：Methods in enzymology

authors： Rico JA,Höcker B

更新日期：2013-01-01 00:00:00

abstract：:Virtually all eukaryotic cell functions and signaling pathways are regulated by protein phosphorylation. The Rad53 kinase plays crucial roles in the DNA damage response in Saccharomyces cerevisiae and is widely used as a surrogate marker for DNA damage checkpoint activation by diverse genotoxic agents. Most currently ...

journal_title：Methods in enzymology

authors： Hoch NC,Chen ES,Tsai MD,Heierhorst J

更新日期：2017-01-01 00:00:00

abstract：:Lymphocytes proliferate in response to several stimuli. In many situations, a rapid lymphocyte expansion, or the identification of a slow dividing cell subpopulation may be of great interest. Thus, it is necessary to perform reliable assays to study and compare lymphocyte subsets proliferation. For this purpose, carbo...

journal_title：Methods in enzymology

authors： Terrén I,Orrantia A,Vitallé J,Zenarruzabeitia O,Borrego F

更新日期：2020-01-01 00:00:00