The cytomegalovirus promoter-driven short hairpin RNA constructs mediate effective RNA interference in zebrafish in vivo

The ability to utilize the RNA interference (RNAi) machinery for silencing target-gene expression has created a lot of excitement in the research community. In the present study, we used a cytomegalovirus (CMV) promoter-driven DNA template approach to induce short hairpin RNA (shRNA) triggered RNAi to block exogenous Enhanced Green Fluorescent Protein (EGFP) and endogenous No Tail (NTL) gene expressions. We constructed three plasmids, pCMV-EGFP-CMV-shGFP-SV40, pCMV-EGFP-CMV-shNTL-SV40, and pCMV-EGFP-CMV-shScrambled-SV40, each containing a CMV promoter driving an EGFP reporter cDNA and DNA coding for one shRNA under the control of another CMV promoter. The three shRNA-generating plasmids and pCMV-EGFP control plasmid were introduced into zebrafish embryos by microinjection. Samples were collected at 48 h after injection. Results were evaluated by phenotype observation and real-time fluorescent quantitative reverse-transcription polymerase chain reaction (Q-PCR). The shGFP-generating plasmid significantly inhibited the EGFP expression viewed under fluorescent microscope and reduced by 70.05 +/- 1.26% of exogenous EGFP gene mRNA levels compared with controls by Q-PCR. The shRNA targeting endogenous NTL gene resulted in obvious NTL phenotype of 30 +/- 4% and decreased the level of their corresponding mRNAs up to 54.52 +/- 2.05% compared with nontargeting control shRNA. These data proved the feasibility of the CMV promoter-driven shRNA expression technique to be used to inhibit exogenous and endogenous gene expressions in zebrafish in vivo.

Isolation and characterization of a rhabdovirus from co-infection of two viruses in mandarin fish

Co-infection of two viruses has been observed in mandarin fish (Siniperca chuatsi), but the two viruses have not been characterized. In this study, a rhabdovirus has been isolated from the co-infected two viruses extracted from the diseased mandarin fish, and its morphological structure and partial biochemical and biophysical characteristics have been observed and analyzed. The isolated rhabdovirus has a typical bullet shape, and is therefore called S. chttatsi rhabdovirus (SCRV). And, the isolated rhabdovirus produced a higher titer (10(8.5) TCID50 ml(-1)) than did the co-infecting viruses (10(6.5) TCID50 ml(-1)). Subsequently, the viral genome RNA was extracted, and used as template to clone the complete nucleoprotein (N) gene by RT-PCR amplification. Cloning and sequencing of the SCRV N protein revealed 42%-31% amino acid identities to that of trout rhabdovirus 903/87 and the rhabdoviruses in genus Vesiculovirus. SDS-PAGE separation of the isolated SCRV and other two rhabdoviruses also revealed obvious polypeptide profile difference. Moreover, the anti-SCRV N protein antibody was prepared, and the anti-SCRV N protein antibody only could recognize the SCRV N protein, whereas no antigenicity was detected in other two rhabdoviruses. The data suggested that the SCRV should be a rhabdovirus member related to the genus Vesiculovirus in the Rhabdoviridae. (c) 2006 Elsevier B.V. All rights reserved.

RecA-mediated, targeted mutagenesis in zebrafish

We have evaluated the efficacy of RecA, a prokaryotic protein involved with homologous recombination, to direct site-specific mutagenesis in zebrafish embryos. For this we coinjected a vector containing a mutated enhanced green fluorescent protein (EGFP) gene plus 236-nucleotide corrective single-stranded DNAs coated with RecA into I-cell zebrafish embryos. Twenty-hours after fertilization, about 5% to 20% of injected embryos showed EGFP expression in I or more cells when RecA-coated corrective DNAs were used, but not when RecA was omitted. Mutated EGFP genes with 1-bp insertions or deletions were inefficiently activated, whereas those with 7-bp insertions were activated about 4-fold more efficiently. RecA-coated template strand had a higher efficiency than its complementary strand in activation of EGFP expression. Prior irradiation of the embryos with UV light enhanced RecA-mediated restoration of gene activity, suggesting that the effects we observed were augmented by one or more factors of zebrafish DNA repair systems.

Planar refractive microlens arrays in indium phosphide and quartz substrates

Large-area concave refractive microlens arrays, or concave template structures, and then the non-refractive-index-gradient type of planar refractive microlens arrays in InP and quartz substrates, are fabricated utilizing the method consisting of conventional UV photolithography, thermal shaping of concave photoresist microlenses, etching with an argon ion beam of large diameter, and filling or growing optical medium structures onto the curved surfaces of preshaped concave templates. Several key conditions for fabricating concave and also planar microlenses are discussed in detail. The concave structures obtained are characterized by scanning electron microscope and surface profile measurements. The far-field optical characteristics of quartz/ZrO2 planar refractive microlens arrays have been acquired experimentally. (c) 2008 Society of Photo-Optical Instrumentation Engineers.

Nonpolar growth and characterization of InN overlayers on vertically oriented GaN nanorods

Nanostructured hexagonal InN overlayers were heteroepitaxially deposited on vertically oriented c-axis GaN nanorods by metal-organic chemical vapor deposition. InN overlayers grown in radial directions are featured by a nonpolar heteroepitaxial growth mode on GaN nanorods, showing a great difference from the conventional InN growth on (0001) c-plane GaN template. The surface of InN overlayers is mainly composed of several specific facets with lower crystallographic indices. The orientation relationship between InN and GaN lattices is found to be [0001](InN) parallel to [0001](GaN) and [1100](InN)parallel to[1100](GaN). A strong photoluminescence of InN nanostructures is observed. (C) 2009 American Institute of Physics. [DOI 10.1063/1.3177347]

Hillocks and hexagonal pits in a thick film grown by HVPE

A GaN film with a thickness of 250 mu m was grown on a GaN/sapphire template in a vertical hydride vapor phase epitaxy (HVPE) reactor. The full-width at half-maximum (FWHM) values of the film were 141 and 498 arcsec for the (0 0 2) and (1 0 2) reflections, respectively. A sharp band-edge emission with a FWHM of 20 meV at 50 K was observed, which corresponded to good crystalline quality of the film. Some almost circular-shaped hillocks located in the spiral growth center were found on the film surface with dimensions of 100 mu m, whose origin was related to screw dislocations and micropipes. Meanwhile, large hexagonal pits also appeared on the film surface, which had six triangular {1 0 (1) over bar 1} facets. The strong emission in the pits was dominated by an impurity-related emission at 377 nm, which could have been a high-concentration oxygen impurity. (c) 2008 Elsevier Ltd. All rights reserved.

Indium mole fraction effect on the structural and optical properties of quaternary AlInGaN epilayers

AlInGaN quaternary epilayers with varying In mole fraction were investigated using triple-axis x-ray diffraction and photoluminescence measurements. The indium compositional fluctuation is enhanced with increasing In mole fraction, whereas the mosaicity of the AlInGaN epilayers is determined through the GaN template quality. Based on the analysis of the temperature dependence of the PL peak position, it is found that the localization effect strengthens with increasing In mole fraction due to the larger fluctuations of the In distribution. Increasing the influence of the localized state results in increasing the emission intensity and FWHM with the In content.

Lateral phase separation in AlGaN grown on GaN with a high-temperature AIN interlayer

The influences of a high-temperature (HT) AlN interlayer (IL) on the phase separation in crack-free AlGaN grown on GaN have been studied. The depth-dependent cathodoluminescence (CL) spectra indicate a relatively uniform Al distribution in the growth direction, but the monochromatic CL images and the CL spectra obtained by line scan measurements reveal a lateral phase separation in AlGaN grown on relatively thick HT-AlN ILs. Moreover, when increasing the thickness of HT-AlN IL, the domain-like distribution of the AlN mole fraction in AlGaN layers is significantly enhanced through a great reduction of the domain size. The morphology of mesa-like small islands separated by V trenches in the HT-AlN IL, and the grain template formed by the coalescence of these islands during the subsequent AlGaN lateral overgrowth, are attributed to be responsible for the formation of domain-like structures in the AlGaN layer. (c) 2005 American Institute of Physics.

On the formation of well-aligned ZnO nanowall networks by catalyst-free thermal evaporation method

Two-dimensional ZnO nanowall networks were grown on ZnO-coated silicon by thermal evaporation at low temperature without catalysts or additives. All of the results from scanning electronic spectroscope, X-ray diffraction and Raman scattering confirmed that the ZnO nanowalls were vertically aligned and c-axis oriented. The room-temperature photoluminescence spectra showed a dominated UV peak at 378 nm, and a much suppressed orange emission centered at similar to 590 nm. This demonstrates fairly good crystal quality and optical properties of the product. A possible three-step, zinc vapor-controlled process was proposed to explain the growth of well-aligned ZnO nanowall networks. The pre-coated ZnO template layer plays a key role during the synthesis process, which guides the growth direction of the synthesized products. (C) 2007 Elsevier B.V. All rights reserved.

Effect of substrate misorientation on the InAs/InAlAs/InP nanostructure morphology and lateral composition modulation in the InAlAs matrix

The authors report the self-organized growth of InAs/InAlAs quantum wires on nominal (001) InP substrate and (001) InP substrates misoriented by 2 degrees, 4 degrees, and 8 degrees towards both [-110] and [110]. The influence of substrate misorientation on the structural and optical properties of these InAs/InAlAs quantum wires is studied by transmission electron microscopy and photoluminescence measurements. Compared with that grown on nominal (001) InP substrate, the density of InAs/InAlAs quantum wires grown on misoriented InP(001) substrates is enhanced. A strong lateral composition modulation effect take place in the InAlAs buffer layers grown on misoriented InP substrates with large off-cut angles (4 degrees and 8 degrees), which induces a nucleation template for the first-period InAs quantum wires and greatly improve the size distribution of InAs quantum wires. InAs/InAlAs quantum wires grown on InP (001) substrate 8 degrees off cut towards [-110] show the best size homogeneity and photoluminescence intensity. (c) 2007 American Institute of Physics.

Optimization of mechanically induced long-period fibre grating transmission spectrum

A new double-layer grating template is designed to reduce the out-of-band loss as much as 1.8dB when the loss of LP03 reaches 10.2 dB. Meanwhile, we propose a method to remove the sidelobes in the transmission spectra by the adjustment of the thickness of pressure plates. The plate-thickness-induced shift of resonant wavelength and the attenuation of loss peak intensity when removing sidelobes can be modified by the fibre distance and contact point on the pressure plates.

Chemical composition and elastic strain in AlInGaN quaternary films

High-quality AlInGaN quaternary layers were grown on c-Al2O3 using a thick GaN template. A full width at half maximum of 0.075 degrees from AlInGaN(0004) rocking curve and a minimum yield of 5.6% from Rutherford backscattering/channelling spectrometry (RBS) prove the AlInGaN layer of a comparable crystalline quality with GaN layers. The chemical compositions (both of Al and In contents) of AlInGaN layers are directly obtained from RBS and elastic recoil detection analysis. The lattice parameters both in perpendicular and parallel directions are deduced from X-ray diffraction. The AlInGaN layer is found to process a compressive strain in parallel direction and a tensile strain in perpendicular direction. (c) 2006 Elsevier B.V. All rights reserved.

Spatial distribution of deep level defects in crack-free AlGaN grown on GaN with a high-temperature AlN interlayer

The deep level luminescence of crack-free Al0.25Ga0.75N layers grown on a GaN template with a high-temperature grown AlN interlayer has been studied using spatially resolved cathodoluminescence (CL) spectroscopy. The CL spectra of Al0.25Ga0.75N grown on a thin AlN interlayer present a deep level aquamarine luminescence (DLAL) band at about 2.6 eV and a deep level violet luminescence (DLVL) band at about 3.17 eV. Cross-section line scan CL measurements on a cleaved sample edge clearly reveal different distributions of DLAL-related and DLVL-related defects in AlGaN along the growth direction. The DLAL band of AlGaN is attributed to evolve from the yellow luminescence band of GaN, and therefore has an analogous origin of a radiative transition between a shallow donor and a deep acceptor. The DLVL band is correlated with defects distributed near the GaN/AlN/AlGaN interfaces. Additionally, the lateral distribution of the intensity of the DLAL band shows a domainlike feature which is accompanied by a lateral phase separation of Al composition. Such a distribution of deep level defects is probably caused by the strain field within the domains. (c) 2006 American Institute of Physics.

Strain evolution in GaN layers grown on high-temperature AlN interlayers

The strain evolution of the GaN layer grown on a high-temperature AlN interlayer with GaN template by metal organic chemical vapor deposition is investigated. It is found that the layer is initially under compressive strain and then gradually relaxes and transforms to under tensile strain with increasing film thickness. The result of the in situ stress analysis is confirmed by x-ray diffraction measurements. Transmission electron microscopy analysis shows that the inclination of edge and mixed threading dislocations rather than the reduction of dislocation density mainly accounts for such a strain evolution. (c) 2006 American Institute of Physics.

Al2O3 : Cr3+ nanotubes synthesized via homogenization precipitation followed by heat treatment

Cr3+-doped NH4Al(OH)(2)CO3 nanotubes, templated by surfactant assemblies, were successfully synthesized via the homogenization precipitation method, and various crystallographic phase Al2O3:Cr3+ nanotubes were also obtained by postannealing at different temperatures. The characteristic R-1, R-2 doublet line transitions of ruby can be observed in the high crystalline alpha-Al2O3 nanotubes calcined at temperatures higher than 1200 degrees C. The results also indicate that the formation mechanism of the tubular nanostructures should result from the self-rolling action of layered compound NH4Al(OH)(2)CO3 under the assistance of the surfactant soft-template. The convenient synthetic procedure, excellent reproducibility, clean reactions, high yield, and fine quality of products in this work make the present route attractive and significant. Aluminum oxide nanotubes with high specific surface area could be used as fabricating nanosized optical devices doped with different elements and stable catalyst supports of metal clusters.