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1.
J Phys Condens Matter ; 31(10): 104002, 2019 Mar 13.
Artículo en Inglés | MEDLINE | ID: mdl-30625452

RESUMEN

We present electrical measurements from In0.75Ga0.25As 1D channel devices with Rashba-type, spin-orbit coupling present in the 2D contact regions. Suppressed backscattering as a result of the time-reversal asymmetry at the 1D channel entrance results in enhanced ballistic transport characteristics with clear quantised conductance plateaus up to 6 × (2e 2/h). Applying DC voltages between the source and drain ohmic contacts and an in-plane magnetic field confirms a ballistic transport picture. For asymmetric patterned gate biasing, a lateral spin-orbit coupling effect is weak. However, the Rashba-type spin-orbit coupling leads to a g-factor in the 1D channel that is reduced in magnitude from the 2D value of 9 to ~6.5 in the lowest subband when the effective Rashba field and the applied magnetic field are perpendicular.

2.
Nat Nanotechnol ; 11(10): 857-860, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27428275

RESUMEN

Obtaining substantial nonlinear effects at the single-photon level is a considerable challenge that holds great potential for quantum optical measurements and information processing. Of the progress that has been made in recent years one of the most promising methods is to scatter coherent light from quantum emitters, imprinting quantum correlations onto the photons. We report effective interactions between photons, controlled by a single semiconductor quantum dot that is weakly coupled to a monolithic cavity. We show that the nonlinearity of a transition modifies the counting statistics of a Poissonian beam, sorting the photons in number. This is used to create strong correlations between detection events and to create polarization-correlated photons from an uncorrelated stream using a single spin. These results pave the way for semiconductor optical switches operated by single quanta of light.

3.
Osteoarthritis Cartilage ; 23(12): 2158-2166, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26067518

RESUMEN

OBJECTIVE: The aim of this work is to establish the human metacarpal as a new whole joint surface early-stage osteoarthritis (OA) model that enables comparisons of articular cartilage and subchondral bone through high resolution contrast-enhanced CT (CECT) imaging, mechanical testing, and biochemical analysis. DESIGN: The fourth metacarpal was obtained from 12 human cadaveric donors and baseline µCT imaging was followed by indentation testing. The samples were then immersed in anionic (Ioxaglate) and cationic (CA4+) iodinated contrast agent solutions followed by CECT. Cartilage GAG content and distribution was measured using the 1,9 dimethylmethylene blue (DMMB) assay and Safranin-O histology staining. Linear regression was performed to compare cartilage and subchondral bone properties. RESULTS: Strong and significant positive correlations were observed between CA4+ CECT attenuation and both GAG content (R(2) = 0.86) and equilibrium modulus (R(2) = 0.84), while correlations using Ioxaglate were insignificant (R(2) ≤ 0.24, P > 0.05). Subchondral bone plate (SBP) thickness negatively and significantly correlated with SBP mineral density (R(2) = 0.49). Cartilage GAG content significantly correlated with several trabecular bone properties, including positive correlations with bone volume fraction (%BV/TV, R(2) = 0.67), trabecular number (Tb.N, R(2) = 0.60), and trabecular thickness (R(2) = 0.42), and negative relationships with structural model index (SMI, R(2) = 0.78) and trabecular spacing (Tb.Sp, R(2) = 0.56). Similarly, equilibrium modulus correlated positively with %BV/TV (R(2) = 0.50), Tb.N (R(2) = 0.59) and negatively with Tb.Sp (R(2) = 0.55) and SMI (R(2) = 0.60). CONCLUSION: This study establishes the human metacarpal as a new early-stage OA model suitable for rapid, high resolution CECT imaging, mechanical testing, and biochemical analysis of the cartilage and subchondral bone, and for examining their inter-relationships.


Asunto(s)
Cartílago Articular/diagnóstico por imagen , Huesos del Metacarpo/diagnóstico por imagen , Articulación Metacarpofalángica/diagnóstico por imagen , Anciano , Anciano de 80 o más Años , Densidad Ósea , Cadáver , Cartílago Articular/metabolismo , Cartílago Articular/patología , Fuerza Compresiva , Medios de Contraste , Femenino , Glicosaminoglicanos/metabolismo , Humanos , Ácido Yoxáglico , Modelos Lineales , Masculino , Huesos del Metacarpo/metabolismo , Huesos del Metacarpo/patología , Articulación Metacarpofalángica/metabolismo , Articulación Metacarpofalángica/patología , Persona de Mediana Edad , Tomografía Computarizada por Rayos X , Microtomografía por Rayos X
4.
Nat Commun ; 5: 3316, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24548976

RESUMEN

Quantum networks can interconnect remote quantum information processors, allowing interaction between different architectures and increasing net computational power. Fibre-optic telecommunications technology offers a practical platform for routing weakly interacting photonic qubits, allowing quantum correlations and entanglement to be established between distant nodes. Although entangled photons have been produced at telecommunications wavelengths using spontaneous parametric downconversion in nonlinear media, as system complexity increases their inherent excess photon generation will become limiting. Here we demonstrate entangled photon pair generation from a semiconductor quantum dot at a telecommunications wavelength. Emitted photons are intrinsically anti-bunched and violate Bell's inequality by 17 standard deviations High-visibility oscillations of the biphoton polarization reveal the time evolution of the emitted state with exceptional clarity, exposing long coherence times. Furthermore, we introduce a method to evaluate the fidelity to a time-evolving Bell state, revealing entanglement between photons emitted up to 5 ns apart, exceeding the exciton lifetime.

5.
Knee ; 17(5): 362-4, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-19897372

RESUMEN

The Low Contact Stress Patellofemoral Arthroplasty (LCS PFA) is a newer design belonging to the second generation of inlay type implants, addressing the problems encountered in the first generation models (Lubinus & Richard's). The cemented mobile bearing metal backed patellar component in this system is "modular"; allowing interchangeable usage with either the trochlear component in a PFA or the femoral component of a total knee arthroplasty, thus obviating the need for patellar revision during conversion of PFA to TKA. The younger active patient with a PFA may exert extreme joint reaction forces on their patellar implant, rendering early loosening of the patellar implant. The endurance of this implant with repeated flexion beyond 90 degrees is also a concern. We describe a series of three unusual mechanical failures associated with this particular design of metal backed patellar component of the unicompartmental LCS patellofemoral arthroplasty.


Asunto(s)
Artroplastia de Reemplazo de Rodilla/métodos , Prótesis de la Rodilla , Articulación Patelofemoral/cirugía , Falla de Prótesis , Adulto , Artroplastia de Reemplazo de Rodilla/instrumentación , Femenino , Humanos , Persona de Mediana Edad , Articulación Patelofemoral/diagnóstico por imagen , Diseño de Prótesis , Radiografía , Rango del Movimiento Articular , Estrés Mecánico , Soporte de Peso
6.
Biochem Biophys Res Commun ; 367(3): 656-62, 2008 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-18179771

RESUMEN

Histone deacetylase (HDAC) inhibitors (HDIs) are documented for their role in activation and/or repression of gene expression. Currently, it is believed that HDAC inhibitors act at the histone level to alter chromatin dynamics through the inactivation of HDACs thereby resulting in histone hyperacetylation and increased transcriptional activation. However, transcriptional repression of gene expression is not explained by this model. Indeed, changes in the acetylation status of discreet lysine residues of histones associated with genes repressed by HDAC inhibitors have not been reported. Therefore, we carried out a systematic investigation of the changes in histone acetylation status at the promoter regions of two genes differentially affected by HDIs to gain a better understanding of how changes in histone acetylation correspond to changes in transcriptional activity.


Asunto(s)
Carcinoma/metabolismo , Inhibidores Enzimáticos/farmacología , Expresión Génica/efectos de los fármacos , Inhibidores de Histona Desacetilasas , Histonas/efectos de los fármacos , Histonas/metabolismo , Acetilación/efectos de los fármacos , Carcinoma/tratamiento farmacológico , Línea Celular Tumoral , Cromatina/química , Cromatina/efectos de los fármacos , Cromatina/metabolismo , Inmunoprecipitación de Cromatina , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Células HT29 , Histonas/química , Humanos , Lisina/metabolismo , Regiones Promotoras Genéticas/efectos de los fármacos , Proteínas Proto-Oncogénicas pp60(c-src)/genética , Transcripción Genética/efectos de los fármacos
7.
Nanotechnology ; 19(34): 345401, 2008 Aug 27.
Artículo en Inglés | MEDLINE | ID: mdl-21730647

RESUMEN

We have fabricated a single-photon emitting diode based on a quantum dot in a micro-pillar cavity. By temperature tuning the dot emission into resonance with the cavity mode we see an enhancement in the collected photon intensity at 40 K. We perform autocorrelation measurements on the electroluminescence at fixed bias, observing photon anti-bunching. Due to the low resistance and capacitance of our device we can inject current pulses shorter than the lifetime of the quantum state, producing single-photon emission with g((2))(0) = 0.17.

9.
Chem Commun (Camb) ; (18): 1862-3, 2001 Sep 21.
Artículo en Inglés | MEDLINE | ID: mdl-12240351

RESUMEN

An ambient temperature liquid transition metal carbonyl anion has been prepared in a metathesis reaction between [bmim]Cl ([bmim]+ = 1-butyl-3-methylimidazolium cation) and Na[Co(CO)4]; the ionic liquid catalyses the debromination of 2-bromoketones.

10.
Biophys J ; 79(1): 479-84, 2000 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10866973

RESUMEN

Bacterial type I restriction/modification systems are capable of performing multiple actions in response to the methylation pattern on their DNA recognition sequences. The enzymes making up these systems serve to protect the bacterial cells against viral infection by binding to their recognition sequences on the invading DNA and degrading it after extensive ATP-driven translocation. DNA cleavage has been thought to occur as the result of a collision between two translocating enzyme complexes. Using atomic force microscopy (AFM), we show here that EcoKI dimerizes rapidly when bound to a plasmid containing two recognition sites for the enzyme. Dimerization proceeds in the absence of ATP and is also seen with an EcoKI mutant (K477R) that is unable to translocate DNA. Only monomers are seen when the enzyme complex binds to a plasmid containing a single recognition site. Based on our results, we propose that the binding of EcoKI to specific DNA target sequences is accompanied by a conformational change that leads rapidly to dimerization. This event is followed by ATP-dependent translocation and cleavage of the DNA.


Asunto(s)
Enzimas de Restricción del ADN/química , Enzimas de Restricción del ADN/ultraestructura , Microscopía de Fuerza Atómica , Dimerización , Escherichia coli/enzimología , Modelos Químicos , Plásmidos/química , Polilisina/química , Conformación Proteica , S-Adenosilmetionina/química
11.
J Cell Biol ; 148(2): 317-24, 2000 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-10648564

RESUMEN

We have developed a cell-free system for regulated exocytosis in the PC12 neuroendocrine cell line. Secretory vesicles were preloaded with acridine orange in intact cells, and the cells were sonicated to produce flat, carrier-supported plasma membrane patches with attached vesicles. Exocytosis resulted in the release of acridine orange which was visible as a disappearance of labeled vesicles and, under optimal conditions, produced light flashes by fluorescence dequenching. Exocytosis in vitro requires cytosol and Ca(2+) at concentrations in the micromolar range, and is sensitive to Tetanus toxin. Imaging of membrane patches at diffraction- limited resolution revealed that 42% of docked granules were released in a Ca(2+)-dependent manner during 1 min of stimulation. Electron microscopy of membrane patches confirmed the presence of dense-core vesicles. Imaging of membrane patches by atomic force microscopy revealed the presence of numerous particles attached to the membrane patches which decreased in number upon stimulation. Thus, exocytotic membrane fusion of single vesicles can be monitored with high temporal and spatial resolution, while providing access to the site of exocytosis for biochemical and molecular tools.


Asunto(s)
Exocitosis/fisiología , Fusión de Membrana/fisiología , Naranja de Acridina/metabolismo , Animales , Calcio/metabolismo , Sistema Libre de Células , Colorantes Fluorescentes/metabolismo , Membranas/fisiología , Membranas/ultraestructura , Microscopía de Fuerza Atómica , Sistemas Neurosecretores/citología , Células PC12 , Ratas
12.
Int J Syst Bacteriol ; 49 Pt 4: 1615-22, 1999 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-10555343

RESUMEN

In an attempt to understand better the micro-organisms involved in anaerobic degradation of aromatic hydrocarbons in the Fe(III)-reducing zone of petroleum-contaminated aquifers, Fe(III)-reducing micro-organisms were isolated from contaminated aquifer material that had been adapted for rapid oxidation of toluene coupled to Fe(III) reduction. One of these organisms, strain H-5T, was enriched and isolated on acetate/Fe(III) medium. Strain H-5T is a Gram-negative strict anaerobe that grows with various simple organic acids such as acetate, propionate, lactate and fumarate as alternative electron donors with Fe(III) as the electron acceptor. In addition, strain H-5T also oxidizes long-chain fatty acids such as palmitate with Fe(III) as the sole electron acceptor. Strain H-5T can also grow by fermentation of citrate or fumarate in the absence of an alternative electron acceptor. The primary end-products of citrate fermentation are acetate and succinate. In addition to various forms of soluble and insoluble Fe(III), strain H-5T grows with nitrate, Mn(IV), fumarate and the humic acid analogue 2,6-anthraquinone disulfonate as alternative electron acceptors. As with other organisms that can oxidize organic compounds completely with the reduction of Fe(III), cell suspensions of strain H-5T have absorbance maxima indicative of a c-type cytochrome(s). It is proposed that strain H-5T represents a novel genus in the Holophaga-Acidobacterium phylum and that it should be named Geothrix fermentans sp. nov., gen. nov.


Asunto(s)
Compuestos Férricos/metabolismo , Sedimentos Geológicos/microbiología , Bacterias Anaerobias Gramnegativas/clasificación , Petróleo/metabolismo , Contaminantes Químicos del Agua/metabolismo , Abastecimiento de Agua , Biodegradación Ambiental , ADN Bacteriano/genética , ADN Ribosómico/genética , Bacterias Anaerobias Gramnegativas/aislamiento & purificación , Bacterias Anaerobias Gramnegativas/metabolismo , Bacterias Anaerobias Gramnegativas/fisiología , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , ARN Ribosómico 16S/genética , Microbiología del Agua
13.
Int J Syst Bacteriol ; 49 Pt 3: 1177-80, 1999 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10425777

RESUMEN

Two strains of dissimilatory arsenate-reducing vibrio-shaped bacteria are assigned to the genus Sulfurospirillum. These two new species, Sulfurospirillum barnesii strain SES-3T and Sulfurospirillum arsenophilum strain MIT-13T, in addition to Sulfurospirillum sp. SM-5, two strains of Sulfurospirillum deleyianum, and Sulfurospirillum arcachonense, form a distinct clade within the epsilon subclass of the Proteobacteria based on 16S rRNA analysis.


Asunto(s)
Arseniatos/metabolismo , Bacterias Gramnegativas/clasificación , Compuestos de Selenio/metabolismo , Técnicas de Tipificación Bacteriana , ADN Ribosómico/química , ADN Ribosómico/genética , Genes de ARNr , Bacterias Gramnegativas/citología , Bacterias Gramnegativas/genética , Bacterias Gramnegativas/fisiología , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , ARN Ribosómico 16S/genética , Ácido Selénico , Análisis de Secuencia de ADN
14.
Microsc Res Tech ; 44(5): 368-77, 1999 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-10090213

RESUMEN

The origin of contrast in atomic force microscopy (AFM) lies in the probe's response to forces between itself and the sample. These forces most commonly result from changes in height as the tip is scanned over the surface, but can also originate in properties inherent in the sample. These have been exploited as further means of contrast and have spawned an array of similar imaging techniques, such as chemical force microscopy, magnetic force microscopy, and frictional force microscopy. All of these techniques use AFM as an extremely sensitive gauge to map forces at discrete sites on the surface. A natural extension of this approach is to map forces in an array, in order to create a force map. AFM can be used in aqueous or fluid environments, thus allowing the exploration of forces in biological systems under physiologically relevant conditions. By immobilizing one half of an interacting pair of proteins onto the tip and the other half onto the substrate, it is possible to investigate the electrostatic and hydrophobic interactions between them. We employed these techniques to examine the interaction between a pair of proteins of known affinity that are involved in exocytosis (NSF and alpha-SNAP) and separately to demonstrate how two-dimensional force mapping can be applied to the nuclear envelope to identify nuclear pore complexes.


Asunto(s)
Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Microscopía de Fuerza Atómica/métodos , Proteínas de Transporte Vesicular , Animales , Células CHO , Proteínas Portadoras/genética , Cricetinae , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de la Membrana/genética , Proteínas Sensibles a N-Etilmaleimida , Membrana Nuclear/metabolismo , Unión Proteica , Proteínas Recombinantes/metabolismo , Proteínas Solubles de Unión al Factor Sensible a la N-Etilmaleimida
17.
Appl Environ Microbiol ; 64(4): 1504-9, 1998 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-9546186

RESUMEN

To evaluate which microorganisms might be responsible for microbial reduction of humic substances in sedimentary environments, humic-reducing bacteria were isolated from a variety of sediment types. These included lake sediments, pristine and contaminated wetland sediments, and marine sediments. In each of the sediment types, all of the humic reducers recovered with acetate as the electron donor and the humic substance analog, 2,6-anthraquinone disulfonate (AQDS), as the electron acceptor were members of the family Geobacteraceae. This was true whether the AQDS-reducing bacteria were enriched prior to isolation on solid media or were recovered from the highest positive dilutions of sediments in liquid media. All of the isolates tested not only conserved energy to support growth from acetate oxidation coupled to AQDS reduction but also could oxidize acetate with highly purified soil humic acids as the sole electron acceptor. All of the isolates tested were also able to grow with Fe(III) serving as the sole electron acceptor. This is consistent with previous studies that have suggested that the capacity for Fe(III) reduction is a common feature of all members of the Geobacteraceae. These studies demonstrate that the potential for microbial humic substance reduction can be found in a wide variety of sediment types and suggest that Geobacteraceae species might be important humic-reducing organisms in sediments.


Asunto(s)
Bacterias Anaerobias Gramnegativas/aislamiento & purificación , Bacterias Anaerobias Gramnegativas/metabolismo , Sustancias Húmicas/metabolismo , Microbiología del Agua , Ácido Acético/metabolismo , Antraquinonas/metabolismo , Secuencia de Bases , Cartilla de ADN/genética , Transporte de Electrón , Agua Dulce/microbiología , Bacterias Anaerobias Gramnegativas/genética , Hierro/metabolismo , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Agua de Mar/microbiología , Bacterias Reductoras del Azufre/genética , Bacterias Reductoras del Azufre/aislamiento & purificación , Bacterias Reductoras del Azufre/metabolismo
18.
Anaerobe ; 4(6): 277-82, 1998 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-16887653

RESUMEN

An isolate, designated GC-29, that could incompletely oxidize glucose to acetate and carbon dioxide with Fe(III) serving as the electron acceptor was recovered from freshwater sediments of the Potomac River, Maryland. This metabolism yielded energy to support cell growth. Strain GC-29 is a facultatively anaerobic, gram-negative motile rod which, in addition to glucose, also used sucrose, lactate, pyruvate, yeast extract, casamino acids or H2 as alternative electron donors for Fe(III) reduction. Stain GC-29 could reduce NO3(-), Mn(IV), U(VI), fumarate, malate, S2O3(2-), and colloidal S0 as well as the humics analog, 2,6-anthraquinone disulfonate. Analysis of the almost complete 16S rRNA sequence indicated that strain GC-29 belongs in the Shewanella genus in the epsilon subdivision of the Proteobacteria. The name Shewanella saccharophilia is proposed. Shewanella saccharophilia differs from previously described fermentative microorganisms that metabolize glucose with the reduction of Fe(III) because it transfers significantly more electron equivalents to Fe(III); acetate and carbon dioxide are the only products of glucose metabolism; energy is conserved from Fe(III) reduction; and glucose is not metabolized in the absence of Fe(III). The metabolism of organisms like S. saccharophilia may account for the fact that glucose is metabolized primarily to acetate and carbon dioxide in a variety of sediments in which Fe(III) reduction is the terminal electron accepting process.

19.
Arch Microbiol ; 168(5): 380-8, 1997 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-9325426

RESUMEN

A newly discovered arsenate-reducing bacterium, strain OREX-4, differed significantly from strains MIT-13 and SES-3, the previously described arsenate-reducing isolates, which grew on nitrate but not on sulfate. In contrast, strain OREX-4 did not respire nitrate but grew on lactate, with either arsenate or sulfate serving as the electron acceptor, and even preferred arsenate. Both arsenate and sulfate reduction were inhibited by molybdate. Strain OREX-4, a gram-positive bacterium with a hexagonal S-layer on its cell wall, metabolized compounds commonly used by sulfate reducers. Scorodite (FeAsO42. H2O) an arsenate-containing mineral, provided micromolar concentrations of arsenate that supported cell growth. Physiologically and phylogenetically, strain OREX-4 was far-removed from strains MIT-13 and SES-3: strain OREX-4 grew on different electron donors and electron acceptors, and fell within the gram-positive group of the Bacteria, whereas MIT-13 and SES-3 fell together in the epsilon-subdivision of the Proteobacteria. Together, these results suggest that organisms spread among diverse bacterial phyla can use arsenate as a terminal electron acceptor, and that dissimilatory arsenate reduction might occur in the sulfidogenic zone at arsenate concentrations of environmental interest. 16S rRNA sequence analysis indicated that strain OREX-4 is a new species of the genus Desulfotomaculum, and accordingly, the name Desulfotomaculum auripigmentum is proposed.


Asunto(s)
Arseniatos/metabolismo , Bacilos Grampositivos Formadores de Endosporas/metabolismo , Sulfatos/metabolismo , Bacterias Reductoras del Azufre/metabolismo , Bacterias Anaerobias/clasificación , Bacterias Anaerobias/aislamiento & purificación , Bacterias Anaerobias/metabolismo , Bacterias Anaerobias/ultraestructura , Biotransformación , Sedimentos Geológicos/microbiología , Bacilos Grampositivos Formadores de Endosporas/clasificación , Bacilos Grampositivos Formadores de Endosporas/aislamiento & purificación , Bacilos Grampositivos Formadores de Endosporas/ultraestructura , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , ARN Ribosómico 16S/genética , Especificidad por Sustrato , Sulfuros/metabolismo , Bacterias Reductoras del Azufre/clasificación , Bacterias Reductoras del Azufre/aislamiento & purificación , Bacterias Reductoras del Azufre/ultraestructura , Microbiología del Agua
20.
J Cell Sci ; 110 ( Pt 20): 2507-18, 1997 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-9372440

RESUMEN

A cDNA encoding Xlamin B1 was cloned from a whole ovary mRNA by RT-PCR. GST-lamin fusion constructs were generated from this cDNA by first creating convenient restriction sites within the Xlamin B1 coding sequence, using PCR directed mutagenesis, and then sub-cloning relevant sequences into pGEX-4T-3. Two expression constructs were made, the first, termed delta 2+ lacked sequences encoding the amino-terminal 'head domain' of lamin B1 but included sequences encoding the nuclear localization signal sequence (NLS). The second expression construct, termed delta 2-, lacked sequences encoding the amino-terminal 'head domain' as well as sequences encoding the NLS. Purified fusion proteins expressed from these constructs, when added to egg extracts prior to sperm pronuclear assembly, formed hetero-oligomers with the endogenous lamin B3. The delta 2+ fusion protein prevented nuclear lamina assembly but not nuclear membrane assembly. The resulting nuclei were small (approximately 10 microns in diameter), did not assemble replication centers and failed to initiate DNA replication. When the delta 2- fusion protein was added to egg extracts prior to sperm pronuclear assembly, lamina assembly was delayed but not prevented. The resulting nuclei although small (approximately 12 microns), did form replication centers and initiated DNA replication. When added to egg extracts after sperm pronuclear assembly was completed delta 2+, but not delta 2-, entered the pre-formed nuclei causing lamina disassembly. However, the disassembly of the lamina by delta 2+ did not result in the disruption of replication centers and indeed these centres remained functional. These results are consistent with the hypothesis that lamina assembly precedes and is required for the formation of replication centers but does not support those centers directly.


Asunto(s)
Replicación del ADN/fisiología , Proteínas de Filamentos Intermediarios , Lamina Tipo B , Proteínas Nucleares/fisiología , Animales , Núcleo Celular/metabolismo , Expresión Génica , Glutatión Transferasa/genética , Filamentos Intermedios/metabolismo , Laminas , Mutagénesis , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Óvulo , Biosíntesis de Péptidos , Proteínas Recombinantes de Fusión/genética , Xenopus
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