Stability and molecular modeling of triplex DNA inhibiting DNA binding protein binding to the core promoter of hepatitis B virus.

Two three-dimensional structure models of the 21nt oligodeoxyribonucleotides, CPI (G3TG-2TGT2G5TG2TGT) and CP3 (TGTG2TGST2GTG2TG3), were constructed by InsightII (MSI) software in IRIS Indigo2 (SGI) workstation using the crystal structure of TAT tripler formation as the template. The initial structures subsequently were minimized by molecular mechanics. The final structures were believed as the dominant conformation. The results showed that the energy of CP1 is lower than that of CP3, and the former is more stable than the latter. Moreover, the results further proved that the 21nt oligodeoxyribo-nucleotide CP1 stably combines with the core promoter (Cp) fragment of hepatitis B virus (HBV) to form a tripler DNA, and CP1 specifically inhibits a specific cellular factor (DNA binding protein) binding to Cp fragment. These results indicated that specific repression of gene transcription of HBV DNA might be possible by tripler-formation DNA.

Genetic Stability of Progeny from an Artificial Allotetraploid Carp Using Sperm from Five Fish Species

Microsatellite markers and D-loop sequences of mtDNA from a female allotetraploid parent carp and her progenies of generations 1 and 2 induced by sperm of five distant fish species were analyzed. Eleven microsatellite markers were used to identify 48 alleles from the allotetraploid female. The same number of alleles (48) appeared in the first and second generations of the gynogenetic offspring, regardless of the source of the sperm used as an activator. The mtDNA D-loop analysis was performed on the female tetraploid parent, 25 gynogenetic offspring, and 5 sperm-donor species. Fourteen variable sites from the 1,018 bp sequences were observed in the offspring as compared to the female tetraploid parent. Results from D-loop sequence and microsatellite marker analysis showed exclusive maternal transmission, and no genetic information was derived from the father. Our study suggests that progenies of artificial tetraploid carp are genetically stable, which is important for genetic breeding of this tetraploid fish.

Hybrid cytomegalovirus-U6 promoter-based plasmid vectors improve efficiency of RNA interference in zebrafish

Short hairpin RNA (shRNA) directed by RNA polymerase III (Pol III) or Pol II promoter was shown to be capable of silencing gene expression, which should permit analyses of gene functions or as a potential therapeutic tool. However, the inhibitory effect of shRNA remains problematic in fish. We demonstrated that silencing efficiency by shRNA produced from the hybrid construct composed of the CMV enhancer or entire CMV promoter placed immediately upstream of a U6 promoter. When tested the exogenous gene, silencing of an enhanced green fluorescent protein (EGFP) target gene was 89.18 +/- 5.06% for CMVE-U6 promoter group and 88.26 +/- 6.46% for CMV-U6 promoter group. To test the hybrid promoters driving shRNA efficiency against an endogenous gene, we used shRNA against no tail (NTL) gene. When vectorized in the zebrafish, the hybrid constructs strongly repressed NTL gene expression. The NTL phenotype occupied 52.09 +/- 3.06% and 51.56 +/- 3.68% for CMVE-U6 promoter and CMV-U6 promoter groups, respectively. The NTL gene expression reduced 82.17 +/- 2.96% for CMVE-U6 promoter group and 83.06 +/- 2.38% for CMV-U6 promoter group. We concluded that the CMV enhancer or entire CMV promoter locating upstream of the U6-promoter could significantly improve inhibitory effect induced by the shRNA for both exogenous and endogenous genes compared with the CMV promoter or U6 promoter alone. In contrast, the two hybrid promoter constructs had similar effects on driving shRNA.

Knock down of gfp and no tail expression in zebrafish embryo by in vivo-transcribed short hairpin RNA with T7 plasmid system

A short-hairpin RNA (shRNA) expression system, based on T7 RNA polymerase (T7RP) directed transcription machinery, has been developed and used to generate a knock down effect in zebrafish embryos by targeting green fluorescent protein (gfp) and no tail (ntl) mRNA. The vector pCMVT7R harboring T7RP driven by CMV promoter was introduced into zebrafish embryos and the germline transmitted transgenic individuals were screened out for subsequent RNAi application. The shRNA transcription vectors pT7shRNA were constructed and validated by in vivo transcription assay. When pT7shGFP vector was injected into the transgenic embryos stably expressing T7RP, gfp relative expression level showed a decrease of 68% by analysis of fluorescence real time RT-PCR. As a control, injection of chemical synthesized siRNA resulted in expression level of 40% lower than the control when the injection dose was as high as 2 mu g/mu l. More importantly, injection of pT7shNTL vector in zebrafish embryos expressing T7RP led to partial absence of endogenous ntl transcripts in 30% of the injected embryos when detected by whole mount in situ hybridization. Herein, the T7 transcription system could be used to drive the expression of shRNA in zebrafish embryos and result in gene knock down effect, suggesting a potential role for its application in RNAi studies in zebrafish embryos.

Broad host range plasmid-based gene transfer system in the cyanobacterium Gloeobacter violaceus which lacks thylakoids

Gloeobacter violaceus, a cyanobacterium lack of thylakoids, is refractory to genetic manipulations because its cells are enveloped by a thick gelatinous sheath and in colonial form. In this study, a large number of single cells were obtained by repeated pumping with a syringe with the gelatinous sheath removed. And an exogenous broad host range plasmid pKT210 was conjugatively transferred into G. violaceus. Analyses with dot-blot hybridization and restriction mapping showed that the exogenous plasmid pKT210 had been introduced into G. violaceus and stably maintained with no alteration in its structure. pKT210 extracted from G. violaceus exconjugants could be transformed into the mcr - mrr - E. coli strain DH10B but not the mcr(+) mrr(+) strain DH5alpha, which suggests that a methylase system may be present in G. violaceus.

Au@Si-n: Growth behavior, stability and electronic structure

The configurations, stability, and electronic structure of AuSin (n = 1-16) clusters have been investigated within the framework of the density functional theory at the B3PW91/LanL2DZ and PW91/DNP levels. The results show that the Au atom begins to occupy the interior site for cages as small as Si-11 and for Si-12 the Au atom completely falls into the interior site forming Au@Si-12 cage. A relatively large embedding energy and small HOMO-LUMO gap are also found for this Au@Si-12 structure indicating enhanced chemical activity and good electronic transfer properties. All these make Au@Si-12 attractive for cluster-assembled materials.

Stability and photoemission characteristics for GaAs photocathodes in a demountable vacuum system

The stability and photoemission characteristics for reflection-mode GaAs photocathodes in a demountable vacuum system have been investigated by using spectral response and x-ray photoelectron spectroscopy measurements at room temperature. We find that the shape of the spectral response curve for the cathode changes with time in the vacuum system, but after applying fresh cesium to the degraded cathode, the spectral response can almost be restored. The change and restoration of curve shape are mainly attributed to the evolution of the surface barrier. We illustrate the evolution and analyze the influence of the barrier on the spectral response of the cathode. (C) 2008 American Institute of Physics.

Shape stability of InAs self-assembled islands on vicinal GaAs(001) substrates

We have grown InAs self-assembled islands on vicinal GaAs( 001) substrates. Atomic force microscopy and photoluminescence studies show that the islands have a clear bimodal size distribution. While most of the small islands whose growth is limited by the width of one multi-atomic step have compact symmetric shapes, a large fraction of the large islands limited by the width of one step plus one terrace have asymmetric shapes which are elongated along the multi-atomic step lines. These results can be attributed to the shape-related energy of the islands at different states of their growth. (C) 2008 Elsevier B. V. All rights reserved.

Theoretical investigation on the phase stability of Nd2Fe14B and site preference of V, Cr, Mn, Zr and Nb

The stability of the excellent permanent magnetic compound Nd2Fe14B and substitution of Fe in the compound by V, Cr, Mn, Zr and Nb are investigated by using interatomic pair potentials which are converted from lattice-inversion method. Calculation shows that the substitution always makes the cell volume larger, and the increase of the volume is almost linear with substituent concentration. The calculated cohesive energy shows that the preferential order of substitution of Fe is Nb, V, Cr, Mn, Zr. Nevertheless, all the five substituting elements should most preferentially replace Fe in the j(2)' site, which has the greatest space among all six Fe sites. (C) 2005 Elsevier B.V. All rights reserved.

Study on the thermal stability of InN by in-situ laser reflectance system

The thermal stability of InN in the growth environment in metalorganic chemical vapor deposition was systematically investigated in situ by laser reflectance system and ex situ by morphology characterization, X-ray diffraction and X-ray photoelectron spectroscopy. It was found that InN can withstand isothermal annealing at temperature as high as 600 degrees C in NH3 ambient. While in N-2 atmosphere, it will decompose quickly to form In-droplets at least at the temperature around 500 degrees C, and the activation energy of InN decomposition was estimated to be 2.1 +/- 0.1 eV. Thermal stability of InN when annealing in NH3 ambient during temperature altering would be very sensitive to ramping rate and NH3 flow rate, and InN would sustain annealing process at small ramping rate and sufficient supply of reactive nitrogen radicals. Whereas In-droplets formation was found to be the most frequently encountered phenomenon concerning InN decomposition, annealing window for conditions free of In-droplets was worked out and possible reasons related are discussed. In addition, InN will decompose in a uniform way in the annealing window, and the decomposition rate was found to be in the range of 50 and 100 nm/h. Hall measurement shows that annealing treatment in such window will improve the electrical properties of InN. (c) 2005 Elsevier B.V. All rights reserved.

Structure, stability and photoelectronic properties of transition films from amorphous to microcrystalline silicon

A series of hydrogenated silicon films near the threshold of crystallinity was prepared by very high frequency plasma enhanced chemical vapor deposition (VHF-PECVD) from a mixture of SiH4 diluted in H, The effect of hydrogen dilution ratios R-H = [H-2]/[SiH4] on microstructure of the films was investigated. Photoelectronic properties and stability of the films were studied as a function of crystalline fraction. The results show that more the crystalline volume fraction in the silicon films, the higher mobility life-time product (mu tau), better the stability and lower the photosensitivity. Those diphasic films contained 8%-31% crystalline volume fraction can gain both the fine photoelectronic properties and high stability. in the diphasic (contained 12% crystalline volume fraction) solar cell, we obtained a much lower light-induced degradation of similar to 2.9%, with a high initial efficiency of 10.01% and a stabilized efficiency of 9.72% (AM1.5, 100 mW/cm(2)). (c) 2005 Elsevier B.V. All rights reserved.

Effects of crystalline quality on the phase stability of cubic boron nitride thin films under medium-energy ion irradiation

To investigate the effect of radiation damage on the stability and the compressive stress of cubic boron nitride (c-BN) thin films, c-BN films with various crystalline qualities prepared by dual beam ion assisted deposition were irradiated at room temperature with 300 keV Ar+ ions over a large fluence range up to 2 x 10(16) cm(-2). Fourier transform infrared spectroscopy (FTIR) data were taken before and after each irradiation step. The results show that the c-BN films with high crystallinity are significantly more resistant against medium-energy bombardment than those of lower crystalline quality. However, even for pure c-BN films without any sp(2)-bonded BN, there is a mechanism present, which causes the transformation from pure c-BN to h-BN or to an amorphous BN phase. Additional high resolution transmission electron microscopy (HRTEM) results support the conclusion from the FTIR data. For c-BN films with thickness smaller than the projected range of the bombarding Ar ions, complete stress relaxation was found for ion fluences approaching 4 x 10(15) cm(-2). This relaxation is accompanied, however, by a significant increase of the width of c-BN FTIR TO-line. This observation points to a build-up of disorder and/or a decreasing average grain size due to the bombardment. (c) 2005 Elsevier B.V. All rights reserved.

Stability of GaAs photocathodes under different intensities of illumination

The photocurrent curves of reflection-mode GaAs photocathodes as a function of time, when were illuminated by white light with an intensity of 0, 33 and 100 Ix, respectively, were measured using a multi-information measurement system. The calculated lifetimes of cathodes are 320, 160 and 75 min, respectively, showing that the stability of cathodes degraded with the increase of light intensity. The lifetime of cathode, illuminated by white light with an intensity of 100 Ix, while no photocurrent was being drawn during the illumination, was 100 min. Through comparison, we found that the influence of illumination on cathodes stability is greater than that of photocurrent. The quantum-yield curves of cathodes as a functions of time, when illuminated by white light with an intensity of 33 Ix, were measured also. The measured results show that the shape of the yield curves changes with increasing illumination time due to the faster quantum-yield degradation rate of low energy photons. Based on the revised quantum-efficiency equations for the reflection-mode cathodes, the variation of yield curves are analyzed to be due to the intervalley diffusion of photoelectrons and the evolution of the surface potential barrier profile of the photocathodes during degradation process.

Nonlinear optical properties and thermal stability of the poled polymer NTAB/PEK-c

Films of high glass' transition temperature polymer polyetherketone doped with chromophore 2,2'[4-[(5-nitro-2-thiazolyl)azophenyl]-amino]-bisethanol NTAB) were prepared, poled by the corona-onset poling setup which includes a grid voltage making the surface-charge distribution uniform at elevated temperature. The thickness of the films was measured by the Model 2010 Prism Coupler system. Second harmonic generation d(33) was measured by the second harmonic generation method, and the d33 is 38.12 pm/V at 1064 nm under the absorption correction. The nonlinear optical activity maintains is 80% of its initial value. (C) 2002 Elsevier Science B.V. All rights reserved.

Side radical influence on the nonlinear optical properties and thermal stability of poled polymer films

Films of polyetherketone doped with the chromophores Disperse Red 1 (DR1) and Disperse Red 13 (DR13) were prepared by spin-coating method. By the in situ Second-harmonic Generation (SHG) signal intensity measurement, the optimal poling temperatures were obtained. For the investigated polyetherketone polymer doped with DR1 (DR1/PEK-c) and polyetherketone polymer doped with DR13 (DR13/PEK-c) films, the optimal poling temperatures were 150degreesC and 140degreesC, respectively. Under the optimal poling conditions, the high second-order nonlinear optical coefficient chi(33)((2)) = 11.02 pm/V has been obtained for the DR1/PEK-c; and for DR13/PEK-c at the same conditions the coefficient is 17.9 pm/V. The SHG signal intensity DR1/PEK-c could maintain more than 80% of its initial value when the temperature was under 100degreesC, and the SHG signal intensity of the DR13/PEK-c could maintain more than 80% of its initial value when the temperature was under 135degreesC. (C) 2002 Kluwer Academic Publishers.