Molecular beacons are oligonucleotide probes that become fluorescent upon hybridization. We designed molecular beacons to detect a point mutation in the methylenetetrahydrofolate reductase (MTHFR) gene, a mutation that has been related to an increased risk for cardiovascular disease and neural tube defects. The application of molecular beacons enables fast, semiautomated, ... More
Sites of tubulin polymerization in PC12 cells.
AuthorsKeith CH, Blane K
JournalJ Neurochem
PubMed ID2179472
'The site at which tubulin enters into polymer in the neuritic process is a very important datum in terms of our understanding of the mechanism of transport of the microtubular cytoskeleton out the axon. If the form of tubulin being transported out the axon is the microtubule, then assembly of ... More
Kinetic studies by fluorescence resonance energy transfer employing a double-labeled oligonucleotide: hybridization to the oligonucleotide complement and to single-stranded DNA.
AuthorsParkhurst KM, Parkhurst LJ
JournalBiochemistry
PubMed ID7819209
'A single 16-base oligodeoxyribonucleotide was labeled at the 3''-end with fluorescein and at the 5''-end with x-rhodamine (R*oligo*F); the chromophores served as a donor/acceptor pair, respectively, for Förster resonance energy transfer. We exploited the striking differences in the steady-state emission spectra of the R*oligo*F as a single strand and in ... More
Microtubules of the kinetochore fiber turn over in metaphase but not in anaphase.
AuthorsGorbsky GJ, Borisy GG
JournalJ Cell Biol
PubMed ID2668301
'In previous work we injected mitotic cells with fluorescent tubulin and photobleached them to mark domains on the spindle microtubules. We concluded that chromosomes move poleward along kinetochore fiber microtubules that remain stationary with respect to the pole while depolymerizing at the kinetochore. In those experiments, bleached zones in anaphase ... More
Modification of the GeneScan 2500 fluorescent dye standard for accurate product sizing.
AuthorsBartlett JM, Crilly A, White A, Madhok R
JournalMol Biotechnol
PubMed ID10934531
'This article describes a procedure for modification of the commercially prepared GeneScan 2500 size standard for allelotyping with large DNA fragments such as variable number tandem repeats (VNTRs). Here a procedure was used to adapt commercially available size standards for the sizing of the interleukin-6 (IL6) 3''VNTR, which has allele ... More
Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.
AuthorsJu J, Ruan C, Fuller CW, Glazer AN, Mathies RA
JournalProc Natl Acad Sci U S A
PubMed ID7753809
'Fluorescent dye-labeled DNA primers have been developed that exploit fluorescence energy transfer (ET) to optimize the absorption and emission properties of the label. These primers carry a fluorescein derivative at the 5'' end as a common donor and other fluorescein and rhodamine derivatives attached to a modified thymidine residue within ... More
How microtubules get fluorescent speckles.
AuthorsWaterman-Storer CM, Salmon ED
JournalBiophys J
PubMed ID9746548
'The dynamics of microtubules in living cells can be seen by fluorescence microscopy when fluorescently labeled tubulin is microinjected into cells, mixing with the cellular tubulin pool and incorporating into microtubules. The subsequent fluorescence distribution along microtubules can appear "speckled" in high-resolution images obtained with a cooled CCD camera (Waterman-Storer ... More
Trajectory of nucleosomal linker DNA studied by fluorescence resonance energy transfer.
AuthorsTóth K, Brun N, Langowski J
JournalBiochemistry
PubMed ID11389607
'While the structure of the nucleosome core is known in atomic detail, the precise geometry of the DNA beyond the core particle is still unknown. We have used fluorescence resonance energy transfer (FRET) for determining the end-to-end distance of DNA fragments assembled with histones into nucleosomes. The DNA of a ... More
Crystallographic and biochemical studies of DivK reveal novel features of an essential response regulator in Caulobacter crescentus.
AuthorsGuillet V, Ohta N, Cabantous S, Newton A, Samama JP
JournalJ Biol Chem
PubMed ID12176983
'DivK is an essential response regulator in the Gram-negative bacterium Caulobacter crescentus and functions in a complex phosphorelay system that precisely controls the sequence of developmental events during the cell division cycle. Structure determinations of this single domain response regulator at different pH values demonstrated that the five-stranded alpha/beta fold ... More
Microtubules in the metaphase-arrested mouse oocyte turn over rapidly.
'After ovulation mammalian oocytes arrest in second meiotic metaphase. We asked whether the microtubules that comprise the meiotic spindle of mouse oocytes were stable or were undergoing rapid cycles of assembly and disassembly. Porcine brain tubulin, derivatized with biotin or x-rhodamine [5- (and -6)-carboxy-x-rhodamine], was microinjected into living oocytes. Biotinylated ... More
Fluorescent speckle microscopy, a method to visualize the dynamics of protein assemblies in living cells.
AuthorsWaterman-Storer CM, Desai A, Bulinski JC, Salmon ED
JournalCurr Biol
PubMed ID9811609
'Fluorescence microscopic visualization of fluorophore-conjugated proteins that have been microinjected or expressed in living cells and have incorporated into cellular structures has yielded much information about protein localization and dynamics [1]. This approach has, however, been limited by high background fluorescence and the difficulty of detecting movement of fluorescent structures ... More
Ultra-high-speed DNA sequencing using capillary electrophoresis chips.
AuthorsWoolley AT, Mathies RA
JournalAnal Chem
PubMed ID8644919
'DNA sequencing has been performed on microfabricated capillary electrophoresis chips. DNA separations were achieved in 50 x 8 microns cross-section channels microfabricated in a 2 in. x 3 in. glass sandwich structure using a denaturing 9% T, 0% C polyacrylamide sieving medium. DNA sequencing fragment ladders were produced and fluorescently ... More
Real-time PCR in the microbiology laboratory.
AuthorsMackay IM
JournalClin Microbiol Infect
PubMed ID15008940
'Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has ... More
Microtubules remodel actomyosin networks in Xenopus egg extracts via two mechanisms of F-actin transport.
'Interactions between microtubules and filamentous actin (F-actin) are crucial for many cellular processes, including cell locomotion and cytokinesis, but are poorly understood. To define the basic principles governing microtubule/F-actin interactions, we used dual-wavelength digital fluorescence and fluorescent speckle microscopy to analyze microtubules and F-actin labeled with spectrally distinct fluorophores in ... More
Donor-acceptor distance distributions in a double-labeled fluorescent oligonucleotide both as a single strand and in duplexes.
AuthorsParkhurst KM, Parkhurst LJ
JournalBiochemistry
PubMed ID7819210
'A 16-mer deoxyribonucleotide was labeled at the 5''-end with x-rhodamine and at the 3''-end with fluorescein. Förster resonance energy transfer was used to determine the distribution, P(R), of donor-acceptor distances for the oligomer in three duplex structures (hybridized to complementary strands having 10, 16, and 24 bases) and as a ... More
Steady state dynamics of intermediate filament networks.
AuthorsVikstrom KL, Lim SS, Goldman RD, Borisy GG
JournalJ Cell Biol
PubMed ID1618899
'We have conducted experiments to examine the dynamic exchange between subunit and polymer of vimentin intermediate filaments (IF) at steady state through the use of xrhodamine-labeled vimentin in fluorescence recovery after photobleaching (FRAP) analysis. The xrhodamine-vimentin incorporated into the endogenous vimentin IF network after microinjection into fibroblasts and could be ... More
A brief survey of methods for preparing protein conjugates with dyes, haptens, and cross-linking reagents.
AuthorsBrinkley M
JournalBioconjug Chem
PubMed ID1616945
Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly.
AuthorsDiamantopoulos GS, Perez F, Goodson HV, Batelier G, Melki R, Kreis TE, Rickard JE
JournalJ Cell Biol
PubMed ID9885247
'CLIP-170 is a cytoplasmic linker protein that localizes to plus ends of microtubules in vivo. In this study, we have characterized the microtubule-binding properties of CLIP-170, to understand the mechanism of its plus end targeting. We show that the NH2-terminal microtubule-interacting domain of CLIP-170 alone localizes to microtubule plus ends ... More
H-type dimer formation of fluorophores: a mechanism for activatable, in vivo optical molecular imaging.
AuthorsOgawa M, Kosaka N, Choyke PL, Kobayashi H,
JournalACS Chem Biol
PubMed ID19480464
'In vivo molecular imaging with target-specific activatable '
Fluorescent microtubules break up under illumination.
AuthorsVigers GP, Coue M, McIntosh JR
JournalJ Cell Biol
PubMed ID3417772
'We have synthesized three new fluorescent analogues of tubulin, using fluorescein or rhodamine groups attached to N-hydroxy-succinimidyl esters, and have partially characterized the properties of these analogues. We have also further characterized the tubulin derivatized with dichlorotriazinyl-aminofluorescein that has previously been used in this and other laboratories. Our results show ... More
Color-blind fluorescence detection for four-color DNA sequencing.
AuthorsLewis EK, Haaland WC, Nguyen F, Heller DA, Allen MJ, MacGregor RR, Berger CS, Willingham B, Burns LA, Scott GB, Kittrell C, Johnson BR, Curl RF, Metzker ML
JournalProc Natl Acad Sci U S A
PubMed ID15800037
'We present an approach called pulsed multiline excitation (PME) for measurements of multicomponent, fluorescence species and demonstrate its application in capillary electrophoresis for DNA sequencing. To fully demonstrate the advantages of PME, a fluorescent dye set has been developed whose absorption maxima span virtually the entire visible spectrum. Unlike emission ... More
Energy-transfer fluorescent reagents for DNA analyses.
AuthorsGlazer AN, Mathies RA
JournalCurr Opin Biotechnol
PubMed ID9013654
'Fluorescence resonance energy transfer has facilitated the development of a new class of high-performance fluorescent labeling reagents for multiplex analyses of nucleic acids. The enhanced emission of energy transfer (ET) primers has provided a decadic improvement in the performance of automated DNA sequencers. The emission spectral purity of ET primers ... More
Preparation of modified tubulins.
AuthorsHyman A, Drechsel D, Kellogg D, Salser S, Sawin K, Steffen P, Wordeman L, Mitchison T
JournalMethods Enzymol
PubMed ID2034137
Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection.
AuthorsXiao W, Stern D, Jain M, Huber CG, Oefner PJ
JournalBiotechniques
PubMed ID11414227
Denaturing high-performance liquid chromatography (DHPLC) is a sensitive, robust, and operationally inexpensive method for the detection of single-base substitutions and small deletions and insertions. To increase sample throughout, we have developed a multiplexing strategy using fluorophores to distinguish different PCR products. The system combines recent advances in the synthesis of ... More
DNA curvature in solution measured by fluorescence resonance energy transfer.
AuthorsTóth K, Sauermann V, Langowski J
JournalBiochemistry
PubMed ID9609713
The sequence-induced curvature of DNA fragments free in solution was characterized by measurements of the end-to-end distance using fluorescence resonance energy transfer (FRET). The 31 bp oligonucleotides were labeled at their 5' ends with fluorescein as the donor and rhodamine X as the acceptor. We compared a curved oligonucleotide with ... More
Typing of the short tandem repeat D8S347 locus with different fluorescence markers.
AuthorsPöltl R, Luckenbach C, Fimmers R, Ritter H
JournalElectrophoresis
PubMed ID9504824
The short tandem repeat (STR) locus D8S347 was analyzed by capillary electrophoresis. Sequencing data and a population study of 203 individuals from a southwestern German population are presented. We detected 12 different alleles, 340-388 bp in length, and found 40 different genotypes. The heterozygosity index was 85.7%. Futhermore, we investigated ... More
Kinesin follows the microtubule's protofilament axis.
AuthorsRay S, Meyhöfer E, Milligan RA, Howard J
JournalJ Cell Biol
PubMed ID8099076
We tested the hypothesis that kinesin moves parallel to the microtubule's protofilament axis. We polymerized microtubules with protofilaments that ran either parallel to the microtubule's long axis or that ran along shallow helical paths around the cylindrical surface of the microtubule. When gliding across a kinesin-coated surface, the former microtubules ... More
Four-color DNA sequencing with 3'-O-modified nucleotide reversible terminators and chemically cleavable fluorescent dideoxynucleotides.
AuthorsGuo J, Xu N, Li Z, Zhang S, Wu J, Kim DH, Sano Marma M, Meng Q, Cao H, Li X, Shi S, Yu L, Kalachikov S, Russo JJ, Turro NJ, Ju J,
JournalProc Natl Acad Sci U S A
PubMed ID18591653
DNA sequencing by synthesis (SBS) on a solid surface during polymerase reaction can decipher many sequences in parallel. We report here a DNA sequencing method that is a hybrid between the Sanger dideoxynucleotide terminating reaction and SBS. In this approach, four nucleotides, modified as reversible terminators by capping the 3'-OH ... More
FRET and competing processes between conjugated polymer and dye substituted DNA strands: a comparative study of probe selection in DNA detection.
AuthorsAl Attar HA, Monkman AP,
JournalBiomacromolecules
PubMed ID19334782
Fluorescence resonance energy transfer (FRET) between water-soluble conjugated polymer, poly-(9,9-bis(6'-N,N,N-trimethylammonium)-hexyl-fluorene phenylene) bromide, and ssDNA's labeled with four different types of dyes (Pacific-blue, Alexa-fluor 430, Fluorescein, and ROX) has been investigated. The effect of spectral overlap and Stokes-shift on the efficiency and properties of FRET were studied. In the DNA sequence ... More
Design, synthesis, and spectroscopic properties of peptide-bridged fluorescence energy-transfer cassettes.
AuthorsLi Y, Glazer AN
JournalBioconjug Chem
PubMed ID10077473
A general partial solid-phase synthetic scheme was developed for the synthesis of energy-transfer cassettes with the donor and acceptor dyes bridged by a peptide. In these cassettes, 6-carboxyfluorescein (Fam) served as a donor. For the second dye, 6-carboxy-X-rhodamine (Rox) was used as a fluorescent acceptor or erythrosin B as a ... More
Homogeneous noncompetitive immunoassay based on the energy transfer between fluorolabeled antibody variable domains (open sandwich fluoroimmunoassay).
AuthorsUeda H, Kubota K, Wang Y, Tsumoto K, Mahoney W, Kumagai I, Nagamune T
JournalBiotechniques
PubMed ID10524316
The antigen-dependent stabilization of an anti-hen egg lysozyme (HEL) antibody HyHEL-10 variable region was monitored with fluorescence resonance energy transfer (FRET) between fluorolabeled heavy chain (VH) and light chain (VL) fragments. The VH and VL fragments labeled with succinimide esters of fluorescein and rhodamine-X, respectively, were mixed in a cooled ... More
Simultaneous binding of two DNA duplexes to the NtrC-enhancer complex studied by two-color fluorescence cross-correlation spectroscopy.
AuthorsRippe K
JournalBiochemistry
PubMed ID10694378
The transcription activator protein NtrC (nitrogen regulatory protein C, also termed NR(I)) can catalyze the transition of Escherichia coli RNA polymerase complexed with the sigma(54) factor (RNAP x sigma(54)) from the closed complex (RNAP x sigma(54) bound at the promoter) to the open complex (melting of the promoter DNA). This ... More
Fluorescence energy transfer detection as a homogeneous DNA diagnostic method.
AuthorsChen X, Zehnbauer B, Gnirke A, Kwok PY
JournalProc Natl Acad Sci U S A
PubMed ID9380706
A homogeneous DNA diagnostic assay based on template-directed primer extension detected by fluorescence resonance energy transfer, named template-directed dye-terminator incorporation (TDI) assay, has been developed for mutation detection and high throughput genome analysis. Here, we report the successful application of the TDI assay to detect mutations in the cystic fibrosis ... More
Detection of specific DNA sequences by fluorescence amplification: a color complementation assay.
AuthorsChehab FF, Kan YW
JournalProc Natl Acad Sci U S A
PubMed ID2594760
We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for ... More
Profluorescent protease substrates: intramolecular dimers described by the exciton model.
AuthorsPackard BZ, Toptygin DD, Komoriya A, Brand L
JournalProc Natl Acad Sci U S A
PubMed ID8876189
Xanthene dyes are known to form dimers with spectral characteristics that have been interpreted in terms of exciton theory. A unique aspect of H-type dimers is the fluorescence quenching that accompanies their formation. Using the principles of exciton theory as a guide, a series of protease substrates was synthesized with ... More
Comparison of fluorescence energy transfer primers with different donor-acceptor dye combinations.
AuthorsHung SC, Mathies RA, Glazer AN
JournalAnal Biochem
PubMed ID9448839
Fluorescence energy transfer (ET) primers are far superior to single dye-labeled primers as labels for DNA sequencing and polymerase chain reaction amplification. We compare here ET primers with different donor and acceptor dye combinations with respect to the relative acceptor fluorescence emission intensity and the amount of residual donor fluorescence ... More
Interaction of the Golgi membranes isolated from rabbit liver with microtubules in vitro.
AuthorsMurata M, Itoh TJ, Kagiwada S, Hishida R, Hotani H, Ohnishi S
JournalBiol Cell
PubMed ID1393149
We have developed a reconstituted model system to study the interaction of the Golgi membranes isolated from rabbit liver with taxol-stabilized bovine-brain microtubules without microtubule-associated proteins (MAPs). The Golgi membranes are associated with microtubules. The sheets of vesicles and the membranous tubules are observed along microtubules by direct visualization using ... More
Differential display with carboxy-X-rhodamine-labeled primers and the selection of differentially amplified cDNA fragments without cloning.
AuthorsYoshikawa Y, Mukai H, Asada K, Hino F, Kato I
JournalAnal Biochem
PubMed ID9466801
Differential display (DD) has been used extensively to detect differentially expressed genes. However, the low reproducibility of displayed bands makes verification by Northern blot difficult and the technique is labor-intensive. This report describes a fluorescent DD with a ROX (carboxy-X-rhodamine)-labeled anchor primer and a revised RT-PCR, utilizing AMV reverse transcriptase, ... More
Do photobleached fluorescent microtubules move?: re-evaluation of fluorescence laser photobleaching both in vitro and in growing Xenopus axon.
AuthorsOkabe S, Hirokawa N
JournalJ Cell Biol
PubMed ID7679673
We previously documented differences in the behavior of microtubules in growing axons of two types of neurons, adult mouse sensory neurons and Xenopus embryonal spinal cord neurons. Namely, the bulk of microtubules was stationary in mouse sensory neurons both by the method of photoactivation of caged-fluorescein-labeled tubulin and photobleaching of ... More
Poleward microtubule flux mitotic spindles assembled in vitro.
AuthorsSawin KE, Mitchison TJ
JournalJ Cell Biol
PubMed ID1999464
In the preceding paper we described pathways of mitotic spindle assembly in cell-free extracts prepared from eggs of Xenopus laevis. Here we demonstrate the poleward flux of microtubules in spindles assembled in vitro, using a photoactivatable fluorescein covalently coupled to tubulin and multi-channel fluorescence videomicroscopy. After local photoactivation of fluorescence ... More
Microtubule treadmilling in vitro investigated by fluorescence speckle and confocal microscopy.
AuthorsGrego S, Cantillana V, Salmon ED
JournalBiophys J
PubMed ID11423395
Whether polarized treadmilling is an intrinsic property of microtubules assembled from pure tubulin has been controversial. We have tested this possibility by imaging the polymerization dynamics of individual microtubules in samples assembled to steady-state in vitro from porcine brain tubulin, using a 2% glycerol buffer to reduce dynamic instability. Fluorescence ... More
Progressive and spatially differentiated stability of microtubules in developing neuronal cells.
AuthorsLim SS, Sammak PJ, Borisy GG
JournalJ Cell Biol
PubMed ID2745551
The establishment of neural circuits requires both stable and plastic properties in the neuronal cytoskeleton. In this study we show that properties of stability and lability reside in microtubules and these are governed by cellular differentiation and intracellular location. After culture for 3, 7, and 14 d in nerve growth ... More
DNA sequencing by capillary electrophoresis: use of a two-laser-two-window intensified diode array detection system.
This paper presents the principles of an instrument designed for DNA sequencing using the standard four-dye-labeled primer approach. The method is based on capillary electrophoresis with laser-induced fluorescence and an intensified diode array detector. An important goal of the instrument design has been a detection system that possesses high sensitivity ... More
Effect of primary and secondary structure of oligodeoxyribonucleotides on the fluorescent properties of conjugated dyes.
We studied fluorescence intensity, polarization and lifetime of some commonly used fluorophores conjugated to oligodeoxyribonucleotides with different primary and secondary structures. We found that fluorescence intensity can increase or decrease upon hybridization of the labeled strand to its complement depending on the sequence and position of the fluorophore. Up to ... More
Fluorescence labeling of bacteria for studies of intracellular pathogenesis.
AuthorsDrevets DA, Elliott AM
JournalJ Immunol Methods
PubMed ID7490459
Interactions between intracellular bacterial pathogens and their eukaryotic cellular hosts or targets are often studied with fluorescence-based techniques such as fluorescence microscopy and flow cytometry. We tested whether the intracellular bacterial pathogens L. monocytogenes, M. avium, M. tuberculosis, and S. typhimurium could be labeled by growth in broth containing the ... More
Influence of fluorophor dye labels on the migration behavior of polymerase chain reaction--amplified short tandem repeats during denaturing capillary electrophoresis.
AuthorsHahn M, Wilhelm J, Pingoud A
JournalElectrophoresis
PubMed ID11545394
The determination of the length of polymerase chain reaction (PCR)-amplified short tandem repeats (STRs) by denaturing capillary electrophoresis (CE) is a standard procedure for purposes of genotyping. We show that dye-specific mobility anomalies exist for 5'-fluorophor-labelled single-stranded DNA (ssDNA) fragments in CE using the performance-optimized polymer 4 (POP4) buffer sieving ... More
Microtubule dynamics and chromosome motion visualized in living anaphase cells.
AuthorsGorbsky GJ, Sammak PJ, Borisy GG
JournalJ Cell Biol
PubMed ID3283149
Chromosome segregation in most animal cells is brought about through two events: the movement of the chromosomes to the poles (anaphase A) and the movement of the poles away from each other (anaphase B). Essential to an understanding of the mechanism of mitosis is information on the relative movements of ... More
Microtubule dynamics in fish melanophores.
AuthorsRodionov VI, Lim SS, Gelfand VI, Borisy GG
JournalJ Cell Biol
PubMed ID8089178
We have studied the dynamics of microtubules in black tetra (Gymnocorymbus ternetzi) melanophores to test the possible correlation of microtubule stability and intracellular particle transport. X-rhodamine-or caged fluorescein-conjugated tubulin were microinjected and visualized by fluorescence digital imaging using a cooled charge coupled device and videomicroscopy. Microtubule dynamics were evaluated by ... More
High-resolution liquid chromatography of fluorescent dye-labeled nucleic acids.
Using 100 mM of triethylammonium acetate as ion-pairing reagent, phosphodiester oligonucleotides labeled fluorescently at their 5' terminus could be separated successfully on alkylated nonporous 2.3-microns poly(styrene-divinylbenzene) particles by means of high-resolution liquid chromatography. Applying excitation wavelengths of 490, 520, 550, and 575 nm, respectively, optimum sensitivity was achieved for the ... More
Actomyosin-based retrograde flow of microtubules in the lamella of migrating epithelial cells influences microtubule dynamic instability and turnover and is associated with microtubule breakage and treadmilling.
AuthorsWaterman-Storer CM, Salmon ED
JournalJ Cell Biol
PubMed ID9334345
We have discovered several novel features exhibited by microtubules (MTs) in migrating newt lung epithelial cells by time-lapse imaging of fluorescently labeled, microinjected tubulin. These cells exhibit leading edge ruffling and retrograde flow in the lamella and lamellipodia. The plus ends of lamella MTs persist in growth perpendicular to the ... More
Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis.
AuthorsJu J, Kheterpal I, Scherer JR, Ruan C, Fuller CW, Glazer AN, Mathies RA
JournalAnal Biochem
PubMed ID8678292
We have designed and synthesized fluorescent oligonucleotide primers having improved fluorescence and electrophoretic properties by exploiting the concept of resonance fluorescence energy transfer (ET). These primers carry a fluorescein derivative at the 5' end as a common fluorescence donor and other fluorescein and rhodamine derivatives attached to a modified thymidine ... More
Zymogen/enzyme discrimination using peptide chloromethyl ketones.
AuthorsWilliams EB, Krishnaswamy S, Mann KG
JournalJ Biol Chem
PubMed ID2708377
Glutamylglycinylarginyl chloromethyl ketone, tyrosylglycinylarginyl chloromethyl ketone, and phenylalanylprolylarginyl chloromethyl ketone have been labeled at their amino termini using fluorescein, rhodamine-X, lissamine-rhodamine, pyrene, and the 1,5-, 2,5-, and 2,6-dimethylaminonaphthalene-1-sulfonyl moieties. These peptidyl chloromethyl ketones have also been modified by incorporation of biotin and epsilon-amino caproyl biotin. The ability of these various ... More
Sequence preferences in cleavage of dsDNA and ssDNA by the extracellular Serratia marcescens endonuclease.
AuthorsMeiss G, Friedhoff P, Hahn M, Gimadutdinow O, Pingoud A
JournalBiochemistry
PubMed ID7547935
The preferred cleavage sites in dsDNA and ssDNA for the extracellular Serratia marcescens endonuclease (commercially available as BENZONASE) were identified by limited digestion of PCR-generated substrates. Two different dsDNA substrates were synthesized by using either radioactively or fluorescent dye labeled primers. ssDNA of identical sequence to one of the fluorescent ... More