Excision of Sleeping Beauty transposons: parameters and applications to gene therapy

A major problem in gene therapy is the determination of the rates at which gene transfer has occurred. Our work has focused on applications of the Sleeping Beauty (SB) transposon system as a non-viral vector for gene therapy. Excision of a transposon from a donor molecule and its integration into a cellular chromosome are catalyzed by SB transposase. In this study, we used a plasmid-based excision assay to study the excision step of transposition. We used the excision assay to evaluate the importance of various sequences that border the sites of excision inside and outside the transposon in order to determine the most active sequences for transposition from a donor plasmid. These findings together with our previous results in transposase binding to the terminal repeats suggest that the sequences in the transposon-junction of SB are involved in steps subsequent to DNA binding but before excision, and that they may have a role in transposase-transposon interaction. We found that SB transposons leave characteristically different footprints at excision sites in different cell types, suggesting that alternative repair machineries operate in concert with transposition. Most importantly, we found that the rates of excision correlate with the rates of transposition. We used this finding to assess transposition in livers of mice that were injected with the SB transposon and transposase. The excision assay appears to be a relatively quick and easy method to optimize protocols for delivery of genes in SB transposons to mammalian chromosomes in living animals. Copyright (C) 2004 John Wiley Sons, Ltd.

HORMONAL REPLACEMENT THERAPY IN FISH - HUMAN GROWTH-HORMONE GENE-FUNCTION IN HYPOPHYSECTOMIZED CARP

Transgenic common carp, Cyprinus carpio, produced by the microinjection of fertilized eggs with a linearized chimeric plasmid pMThGH, a human growth hormone (hGH) gene with a mouse metallothionein-I (MT) gene promoter in pBR322, were used to produce F1 and F2 transgenics. Following hypophysectomy of the transgenic F2 common carp, non-transgenic common carp and non-transgenic crucian carp, growth was monitored for up to 110 days. In addition, recombinant hGH was injected subcutaenously into a group of the non-transgenic crucian carp. Growth rate analyses indicated that (1) hypophysectomy of non-transgenic common carp and crucian carp results in the cessation of growth, (2) hGH administration can stimulate the growth of hypophysectomized crucian carp and (3) hypophysectomized hGH-transgenic common carp continue to grow in the absence of their own growth hormone, suggesting that the hGH-transgene is being expressed in tissues other than the pituitary.

Carrier capture by threading dislocations in (In,Ga)N/GaN heteroepitaxial layers

Using spatially resolved cathodoluminescence spectroscopy, we investigate the spatial luminescence distribution in a fully strained (In,Ga)N layer, in particular, its correlation with the distribution of threading dislocations (TDs). Regarding the impact of TDs on the luminescence properties, we can clearly distinguish between pure edge-type TDs and TDs with screw component. At the positions of both types of TDs, we establish nonradiative recombination sinks. The radius for carrier capture is at least four times larger for TDs with screw component as for pure edge-type TDs. The large capture radius of the former is due to a spiral-like growth mode resulting in an increase in the In content in the center of the spiral domains in comparison to their periphery.

Neutron irradiation effect in two-dimensional electron gas of AlGaN/GaN heterostructures

AlGaN/GaN heterostructures have been irradiated by neutrons with different influences and characterized by means of temperature-dependent Hall measurements and Micro-Raman scattering techniques. It is found that the carrier mobility of two-dimensional electron gas (2DEG) is very sensitive to neutrons. At a low influence of 6.13 x 10(15) cm(-2), the carrier mobility drops sharply, while the sheet carrier density remains the same as that of an unirradiated sample. Moreover, even for a fluence of up to 3.66 x 10(16) cm(-2), the sheet carrier density shows only a slight drop. We attribute the degradation of the figure-of-merit (product of n(s) x mu) of 2DEG to the defects induced by neutron irradiation. Raman measurements show that neutron irradiation does not yield obvious change to the strain state of AlGaN/GaN heterostructures, which proves that degradation of sheet carrier density has no relation to strain relaxation in the present study. The increase of the product of n(s) x mu of 2DEG during rapid thermal annealing processes at relatively high temperature has been attributed to the activation of Ge-Ga transmuted from Ga and the recovery of displaced defects.

Hydrogen related defects in neutron-irradiated silicon grown in hydrogen ambient

Isochronal thermal-annealing behavior of NTD floating-zone silicon grown in hydrogen ambient (called NTD FZ(H) Si) is presented. The dependencies of resistivity and carrier mobility on annealing temperature are determined by room-temperature Hall electrical measurements. Using infrared absorption spectroscopy, hydrogen-related infrared absorption bands evolution for NTD FZ(H) Si were measured in detail. It is demonstrated that compared with NTD FZ(Ar) Si, NTD FZ(H) Si exhibits the striking features upon isochronal annealing in temperature range of 150 similar to 650 degreesC: there appears the formation of an excessive shallow donor at annealing temperature of 500 degreesC. It is shown that the annealing behavior is directly related to the reaction of hydrogen and irradiation-induced defects. The evolution of infrared absorption bands upon temperature reflects a series of complex reaction process: irradiation-induced defects decomposition, breaking of Si-H bonds, migration and aggregation of atomic hydrogen, and formation of the secondary defects. (C) 2002 Elsevier Science B.V. All rights reserved.

Detection of efficient carrier capture in ultrathin InAs/GaAs layers using a degenerate pump-probe technique

By analysing the carrier dynamics based on the rate equations and the change of the refractive index due to the efficient carrier capture, we have calculated the carrier capture process in the InAs/GaAs system detected by a simple degenerate pump-probe technique. The calculated results are found to be in good agreement with the experimental findings. Our results indicate that this simple technique, with the clear advantage of being easy to carry out, can be very useful in studying the carrier dynamics for some specific structures such as InAs ultrathin layers embedded in a GaAs matrix described here.

Study of rapid carrier capture and relaxation in InAs/GaAs heterostructures

The rapid carrier capture and relaxation processes in InAs/GaAs quantum dots were studied at 77K by using a simple degenerate pump-probe technique. A rising process was observed in the transient reflectivity, following the initial fast relaxation associated with GaAs bulk matrix, and this rising process was assigned to be related to the carrier capture from the GaAs barriers to InAs layers. The assignment was modeled using Kramers-Kronig relation. By analyzing the rising process observed in the transient reflectivity, the carrier capture time constants were obtained. The measured capture times decrease with the increase of carrier concentration.

Carrier capture into InAs/GaAs quantum dots detected by a simple degenerate pump-probe technique

We demonstrate that the carrier capture and relaxation processes in InAs/GaAs quantum dots can be detected by a simple degenerate pump-probe technique. We have observed a rising process in the transient reflectivity, following the initial fast relaxation in a GaAs matrix, and assigned this rising process to the carrier capture from the GaAs barriers to the InAs layers. The assignment was modeled using the Kramers-Kronig relations. The capture time was found to depend strongly on the InAs layer thickness as well as on the excitation density and photon energy. (C) 2000 Elsevier Science Ltd. All rights reserved.

Hole capture barrier of self-organized InAs quantum dots

Deep level transient spectroscopy (DLTS) technique was successfully applied to characterize the electric properties of p type self-organized InAs quantum dots. The ground state energy and capture barrier energy of hole of quantum dots were measured for the first time. The energy of ground state of 2.5ML InAs quantum dots with respect to the valence band of bulk GaAs was obtained being about 0.09eV, and there was a barrier associated to the change of charge state of quantum dots. The capture barrier energy of such dots for hole was about 0.26eV. The work is very meaningful for further understanding the intrinsic properties of quantum dots.

Ultrafast low-temperature grown AlGaAs/GaAs photorefractive quantum wells using point defects as capture centers

At a medium substrate temperature of 400 degrees C and a lower As flux, we have grown an ultrafast AlGaAs/GaAs photorefractive multiple quantum well (MQW) structure by molecular beam epitaxy. The as-grown sample exhibits strong photorefractive effect under the transverse Frantz-Keldysh geometry. A peak electroabsorption of 2100 cm(-1) is measured in the as-grown sample in an 11 kV/cm dc electric field, and the peak photorefractive diffraction efficiency can be 1.2%. After postgrowth annealing, the photorefractive effect becomes weak and disappears in samples annealed above 700 degrees C. Using optical transient current spectroscopy, deep levels are measured in these samples. It is found that deep levels are stable against annealing until 700 degrees C. Using a pump-probe technique, carrier lifetimes are measured at room temperature. We find that the as-grown sample has a lifetime of 20 ps, while the 700 degrees C annealed sample has a lifetime of more than 200 ps. The ultrafast lifetime in the as-grown sample is caused by point defects, not by As clusters. Our result show that AlGaAs/GaAs MQW structure grown around 400 degrees C has better performance of the photorefractive effect. (C) 1999 American Institute of Physics. [S0003-6951(99)04036-X].

Improved neutron radiation hardness for Si detectors: Application of low resistivity starting material and or manipulation of N-eff by selective filling of radiation-induced traps at low temperatures

Radiation-induced electrical changes in both space charge region (SCR) of Si detectors and bulk material (BM) have been studied for samples of diodes and resistors made on Si materials with different initial resistivities. The space charge sign inversion fluence (Phi(inv)) has been found to increase linearly with the initial doping concentration (the reciprocal of the resistivity), which gives improved radiation hardness to Si detectors fabricated from low resistivity material. The resistivity of the BM, on the other hand, has been observed to increase with the neutron fluence and approach a saturation value in the order of hundreds k Omega cm at high fluences, independent of the initial resistivity and material type. However, the fluence (Phi(s)), at which the resistivity saturation starts, increases with the initial doping concentrations and the value of Phi(s) is in the same order of that of Phi(inv) for all resistivity samples. Improved radiation hardness can also be achieved by the manipulation of the space charge concentration (N-eff) in SCR, by selective filling and/or freezing at cryogenic temperatures the charge state of radiation-induced traps, to values that will give a much smaller full depletion voltage. Models have been proposed to explain the experimental data.

First charge collection and position-precision data on the medium-resistivity silicon strip detectors before and after neutron irradiation up to 2x10(14) n/cm(2)

Test strip detectors of 125 mu m, 500 mu m, and 1 mm pitches with about 1 cm(2) areas have been made on medium-resistivity silicon wafers (1.3 and 2.7 k Ohm cm). Detectors of 500 mu m pitch have been tested for charge collection and position precision before and after neutron irradiation (up to 2 x 10(14) n/cm(2)) using 820 and 1030 nm laser lights with different beam-spot sizes. It has been found that for a bias of 250 V a strip detector made of 1.3 k Ohm cm (300 mu m thick) can be fully depleted before and after an irradiation of 2 x 10(14) n/cm(2). For a 500 mu m pitch strip detector made of 2.7 k Ohm cm tested with an 1030 nm laser light with 200 mu m spot size, the position reconstruction error is about 14 mu m before irradiation, and 17 mu m after about 1.7 x 10(13) n/cm(2) irradiation. We demonstrated in this work that medium resistivity silicon strip detectors can work just as well as the traditional high-resistivity ones, but with higher radiation tolerance. We also tested charge sharing and position reconstruction using a 1030 nm wavelength (300 mu m absorption length in Si at RT) laser, which provides a simulation of MIP particles in high-physics experiments in terms of charge collection and position reconstruction, (C) 1999 Elsevier Science B.V. All rights reserved.

Neutron irradiation-infrared based measurement method for interstitial oxygen in heavily boron-doped silicon

A novel method, based on an infrared absorption and neutron irradiation technique, has been developed for the determination of interstitial oxygen in heavily boron-doped silicon. The new procedure utilizes fast neutron irradiated silicon wafer specimens. On fast neutron irradiation, the free carriers of high concentration in silicon can be trapped by the irradiated defects and the resistivity increased. The resulting calibration curve for the measurement of interstitial oxygen in boron-doped silicon has been established on the basis of the annealing behaviour of irradiated boron-doped CZ silicon.