The onset of foreign gene transcription in nuclear-transferred embryos of fish

The transcriptional onset of hGH-transgene in fish was studied in the following three cases: the first is in MThGH-transgenic F-4 common carp (Cyprinus carpio) embryos, the second is in nuclear-transferred embryos supported by the transgenic F-4 embryonic nuclei, and the third is in nuclear-transferred embryos supported by the transgenic F-4 tail-fin nuclei. RT-PCR results show that the hGH-transgene initiates its transcriptional activity from early-gastrula stage, the early blastula stage and even 16-cell stage in the first, second and third cases, respectively. it looks like that fish egg cytoplasm could just offer a very restricted reprogramming on transcriptional activity of specific gene in differentiated cell nuclei by nuclear transplantation.

Nuclear transplantation with early-embryonic cells of transgenic fish

Like other transgenic animals, transgenic fishes produced by microinjection are transgenic mosaics. In order to produce homogenous transgenic fish, the transgenic blastula or gastrula cells were dissociated from Carassius auratus, Pengze var, and Cyprinus carpio, Huanghe var., and the nuclei were transferred into the mature eggs of the same species via microinjection or electro-fusion. Five nuclear-transferred Carassius auratus, Pengze var. and one Cyprinus carpio, Huanghe var. were obtained and the existence of the transgene was detected. The possibility of generating homogenous strain of transgenic fish by nuclear transplantation with transgenic early-embryonic cells is discussed.

Embryonic and genetic manipulation in fish

Fishes, the biggest and most diverse community in vertebrates are good experimental models for studies of cell and developmental biology by many favorable characteristics. Nuclear transplantation in fish has been thoroughly studied in China since 1960s. Fish nuclei of embryonic cells from different genera were transplanted into enucleated eggs generating nucleo-cytoplasmic hybrids of adults. Most importantly, nuclei of cultured goldfish kidney cells had been reprogrammed in enucleated eggs to support embryogenesis and ontogenesis of a fertile fish. This was the first case of cloned fish with somatic cells. Based on the technique of microinjection, recombinant MThGH gene has been transferred into fish eggs and the first batch of transgenic fish were produced in 1984. The behavior of foreign gene was characterized and the onset of the foreign gene replication occurred between the blastula to gastrula stages and random integration mainly occurred at later stages of embryogenesis. This eventually led to the transgenic mosaicism. The MThGH-transferred common carp enhanced growth rate by 2-4 times in the founder juveniles and doubled the body weight in the adults. The transgenic common carp were more efficient in utilizing dietary protein than the controls. An "all-fish" gene construct CAgcGH has been made by splicing the common carp beta-actin gene (CA) promoter onto the grass carp growth hormone gene (gcGH) coding sequence. The CAgcGH-transferred Yellow River Carp have also shown significantly fast-growth trait. Combination of techniques of fish cell culture, gene transformation with cultured cells and nuclear transplantation should be able to generate homogeneous strain of valuable transgenic fish to fulfil human requirement in 21(st) century.

Kinetically controlled InN nucleation on GaN templates by metalorganic chemical vapour deposition

This paper presents a study on the nucleation and initial growth kinetics of InN on GaN, especially their dependence on metalorganic chemical vapour deposition conditions. It is found that the density and size of separated InN nano-scale islands can be adjusted and well controlled by changing the V/III ratio and growth temperature. InN nuclei density increases for several orders of magnitude with decreasing growth temperature between 525 and 375 degrees C. At lower growth temperatures, InN thin films take the form of small and closely packed islands with diameters less than 100 nm, whereas at elevated temperatures the InN islands grow larger and become well separated, approaching an equilibrium hexagonal shape due to enhanced surface diffusion of adatoms. The temperature dependence of InN island density gives two activation energies of InN nucleation behaviour, which is attributed to two different kinetic processes related to In adatom surface diffusion and desorption, respectively.

The influence of growth temperature and input V/III ratio on the initial nucleation and material properties of InN on GaN by MOCVD

The effects of growth temperature and V/III ratio on the InN initial nucleation of islands on the GaN (0 0 0 1) surface were investigated. It is found that InN nuclei density increases with decreasing growth temperature between 375 and 525 degrees C. At lower growth temperatures, InN thin films take the form of small and closely packed islands with diameters of less than 100 nm, whereas at elevated temperatures the InN islands can grow larger and well separated, approaching an equilibrium hexagonal shape due to enhanced surface diffusion of adatoms. At a given growth temperature of 500 degrees C, a controllable density and size of separated InN islands can be achieved by adjusting the V/III ratio. The larger islands lead to fewer defects when they are coalesced. Comparatively, the electrical properties of the films grown under higher V/III ratio are improved.

Spin Relaxation of Electrons in Single InAs Quantum Dots

By using polarization-resolved photoluminescence spectra, we study the electron spin relaxation in single InAs quantum dots (QDs) with the configuration of positively charged excitons X+ (one electron, two holes). The spin relaxation rate of the hot electrons increases with the increasing energy of exciting photons. For electrons localized in QDs the spin relaxation is induced by hyperfine interaction with the nuclei. A rapid decrease of polarization degree with increasing temperature suggests that the spin relaxation mechanisms are mainly changed from the hyperfine interaction with nuclei into an electron-hole exchange interaction.

Surface morphology of AlN buffer layer and its effect on GaN growth by metalorganic chemical vapor deposition

The in situ optical reflectivity measurements are employed to monitor the GaN epilayer growth process above low-temperature AlN buffer layer on c-plane sapphire substrate by metalorganic chemical vapor deposition. It is found that the lateral growth of GaN islands and their coalescence is promoted in the initial growth stage if the AlN buffer layer is treated with a long annealing time and has an optimal thickness: As confirmed by atomic force microscopy observations, the quality of GaN epilayers is closely dependent on the surface morphology of AlN buffer layer, especially the grain size and nuclei density after the annealing treatment. (C) 2004 American Institute of Physics.

Evolution of mosaic structure in InN grown by metalorganic chemical vapor deposition

Mosaic structure in InN layers grown by metalorganic chemical vapor deposition at various temperatures has been investigated by X-ray diffraction (XRD). With a combination of Williamson-Hall measurement and fitting of twist angles, it was found that variation of growth temperature from 450 to 550 degrees C leads to the variation of the lateral coherence length, vertical coherence length, tilt and twist of mosaic blocks in InN films in a, respectively, monotonic way. In particular, mosaic tilt increases whereas mosaic twist decreases with elevating temperature. Atomic force microscopy shows the morphological difference of the InN nucleation layers grown at 450 and 550 degrees C. Different coalescence thickness and temperature-dependent in-plane rotation of InN nuclei are considered to account for the XRD results. (c) 2006 Elsevier B.V. All rights reserved.

Influence of growth pressure of a GaN buffer layer on the properties of MOCVD GaN

The influence of growth pressure of GaN buffer layer on the properties of MOCVD GaN on alpha-Al2O3 has been investigated with the aid of a home-made in situ laser reflectometry measurement system. The results obtained with in situ measurements and scanning electron microscope show that with the increase in deposition pressure of buffer layer, the nuclei increase in size, which roughens the surface, and delays the coalescence of GaN nuclei. The optical and crystalline quality of GaN epilayer was improved when buffer layer was deposited at high pressure.

Effects of reactor pressure on GaN nucleation layers and subsequent GaN epilayers grown on sapphire substrate

The influence of reactor pressure on GaN nucleation layer (NL) and the quality of subsequent GaN on sapphire is studied. The layers were grown by low-pressure metalorganic chemical vapor deposition (MOCVD) on c-plane sapphire substrates and investigated by in situ laser reflectometry, atomic force microscope, scanning electron microscope, X-ray diffraction and photoluminescence. With the increase of reactor pressure prior to high-temperature GaN growth, the size of GaN nuclei formed after annealing decreases, the spacing between nucleation sites increases and the coalescence of GaN nuclei is deferred. The optical and crystalline qualities of GaN epilayer were improved when NLs were deposited at high pressure. The elongated lateral overgrowth of GaN islands is responsible for the quality improvement. (C) 2003 Elsevier Science B.V. All rights reserved.

Indium doping effect on GaN in the initial growth stage

Three minutes' growth was carried out to investigate the indium-doping effect on initially grown GaN. Indium-doped and undoped samples were grown by low-pressure metalorganic vapor phase epitaxy. Atomic force microscope observation revealed that In-doping modified the morphology of the nuclei. Indium-doping also enhanced wetting between the buffer and nuclei layers, which was also supported by optical transmission. Photoluminescence suggested that indium-doping obviously enhanced band-edge related emission even in the nucleation stage. X-ray diffraction performed on samples grown for 20 minutes indicated improvement of the crystalline quality through indium-doping. The mechanism of the indium-doping effect was discussed.

Metal-free growth of Si/SiO2 nanowires by annealing SiOx (x < 2) films deposited by PECVD

A new metal catalysis-free method of fabricating Si or SiO2 nanowires (NWs) compatible with Si CMOS technology was proposed by annealing SiOx (x < 2) films deposited by plasma -enhanced chemical vapor deposition (PECVD). The effects of the Si content (x value) and thickness of SiOx films, the annealing process and flowing gas ambient on the NW growth were studied in detail. The results indicated that the SiOx film of a thickness below 300 rim with x value close to 1 was most favorable for NW growth upon annealing at 1000-1150 degrees C in the flowing gas mixture of N-2 and H-2. NWs of 50-100nm in diameter and tens of micrometers in length were synthesized by this method. The formation mechanism was likely to be related to a new type of oxide assisted growth (OAG) mechanism, with Si nanoclusters in SiOx films after phase separation serving as the nuclei for the growth of NWs in SiOx films > 200nm, and SiO molecules from thin SiO, film decomposition inducing the NW growth in films < 100nm. An effective preliminary method to control NW growth direction was also demonstrated by etching trenches in SiOx films followed by annealing.

Bending of jets in the QSO NRAO 530

We present radio images of NRAO 530 on scales ranging from pc to kpc. The observations include the EVN at 5 GHz, the VLBA at 1.6, 8.6 and 15 GHz, the MERLIN at 1.6 and 5 GHz, and the VLA at 5, 8.4, 15, 22, and 43 GHz. The VLBI images show a core-jet structure with an oscillating trajectory on a scale of about 30 mas north of the strongest compact component (core). Superluminal motions are detected in five of the jet components with apparent velocities in the range of 13.6 to 25.2c. A new component is detected at 15 GHz with the VLBA observations, which appears to be associated with the outburst in 2002. Significant polarized emission is detected around the core with the VLBA observations at 15 GHz. Rapid variations of the polarization intensity and angle are found between the epochs in 2002 and 2004. On the kpc-scale, a distant component (labelled as WL) located 11 aresec west (PA=-86 degrees) of the core is detected beyond the core-jet structure which extended to several hundreds of mas in the north-west direction (-50 degrees). A significant emission between the core-jet structure and the WL is revealed. A clump of diffuse emission (labelled EL, 12 arcsec long) at PA 70 degrees to the core, is also detected in the VLA observations, suggesting the presence of double lobes in the source. The core component shows a flat spectrum, while the distant components WL and EL have steep spectra. The steep spectra of the distant components and the detection of the arched emission suggest that the distant components are lobes or hot-spots powered by the core of NRAO 530. The morphologies from pc- to kpc-scales and the bending of jets are investigated. The observed radio morphology from pc to kcp appears to favor the model in which precession or wobbling of the nuclear disk drives the helical motion of the radio plasma and produces the S-shaped structure on kpc scale.

贾第虫的“双等核”及其保持等同机制的研究

贾第虫是一类寄生于肠道的单细胞原生动物，也被认为是目前已知的最原始的真核细胞。它的一个十分奇特和令人感兴趣的特点是：具有在形态和大小上都很相似的左右对称的两个细胞核。已有一些证据表明贾第虫的这样的两个核在其它一些方面也都很相似甚至完全相同，如两核的DNA含量相等，两核都含有rDNA和至少一套染色体，并且在DNA复制、转录和核分裂等功能活动方面也基本都是同步的。但是，这两个核在基因的组成方面是否完全一致，甚至两核之间的对应基因（等位基因）的序列是否完全相同呢？若是，贾第虫为什么需要这样两个完全“等价”的细胞核（“双等核”）？这两个核又是如何长期保持一致而不会因两个核内发生不同的基因变异而产生差异呢？此外，目前普遍认为贾第虫至少是四倍体，即每个核至少为二倍体。那么同一个核内的等位基因是否序列也一致？保持其一致的机制又是什么？这些问题不仅饶有趣味，而且对于揭示贾第虫特殊的遗传机制乃至真核细胞基因组倍性的起源进化等问题具有重要意义。 本文首先对其两个核内是否有相同的基因组成进行了检验。我们选择了三个执行不同功能蛋白的基因为代表以检验贾第虫的两个细胞核是否都含有这些基因。这三个基因分别是：DNA拓扑异构酶II基因（topII）, 核仁蛋白KRR1基因（krr1）和目前贾第虫中报道的极少数含有内含子的基因之一的铁硫蛋白基因（fes）。利用荧光原位杂交(FISH)技术在两个核中进行了定位分析。结果表明这几个代表性的基因在贾第虫的两个细胞核中都同时存在。这提示着贾第虫的两核中具有相同的基因组成，也表明贾第虫的这两个核可能在功能方面也是“等价”的。 其次，我们对其两个核中的等位基因是否具有一致的序列进行了检验，并对其两核之间以及同一核内的等位基因保持一致的机制进行了研究。选择前人文献报道的存在多态位点（即这些位点可能是易变位点）的两个基因：fen1和pdi为研究对象，在自己建立的一个贾第虫克隆培养系上进行单细胞PCR和测序的跟踪分析。跟踪分析了18个月（约分裂1600代）后，检测到如下结果：尽管在fen1基因上尚未发现位点变异的规律，但在pdi基因上发现存在几个位点会间歇出现套峰。据此我们推测套峰的出现可能是由于其中一个等位基因发生了突变，而后套峰消失则可能是突变被恢复。这种等位基因的修复机制很可能是基因转换（gene conversion）。进而我们在贾第虫基因组中找到了参与基因转换的很多酶基因的同源基因，RT-PCR的结果也表明这些基因在贾第虫中是活跃转录的。这进一步提示了贾第虫中发生基因转换的可能性。因此以上结果不仅表明贾第虫的两核之间和同一核内的等位基因的序列都是一致的，同时还说明基因转换可能是同一核内等位基因保持一致的机制。至于两个核之间保持等位基因序列一致的机制，我们采用了两种染核的方法对大量的贾第虫细胞进行了观察，以期能从中找到两核保持等同的证据。结果发现在大量细胞群体中存在一定比例的单核细胞和一侧含有两个核的细胞。据此我们推测：贾第虫可能通过发生一侧细胞核的丢失或萎缩，然后由另外一侧的核进行复制，经过核的重排恢复正常的左右核的状态。这可能就是贾第虫消除两核基因出现的差异，保持两核之间等位基因一致乃至两个核完全等同的机制。

Effects of oriented substrates on cell morphology, the cell cycle, and the cytoskeleton in Ros 17/2.8 cells

Absence of gravity or microgravity influences the cellular functions of bone forming osteoblasts. The underlying mechanism, however, of cellular sensing and responding to the gravity vector is poorly understood. This work quantified the impact of vector-directional gravity on the biological responses of Ros 17/2.8 cells grown on upward-, downward- or edge-on-oriented substrates. Cell morphology and nuclear translocation, cell proliferation and the cell cycle, and cytoskeletal reorganization were found to vary significantly in the three orientations. All of the responses were duration-dependent. These results provide a new insight into understanding how osteoblasts respond to static vector-directional gravity.