Development of a positive-negative selection procedure for gene targeting in fish cells

The gene targeting technique is a powerful tool for analyzing functions of cloned genes and for generating transgenic animals with site-directed integration of foreign genes. In order to develop this technique in fish, positive-negative selection (PNS) and homologous recombination vectors were constructed, and their expression was examined in fish cells. A vector (pNK) for PNS consists of the neomycin resistance gene (neo) as a positive selectable marker gene and the herpes simplex virus (HSV) thymidine kinase (tk) gene as a negative selectable marker gene. Positive selection with geneticin (G418) of epithelioma papulosum of carp (EPC) cells transfected with linearized pNK vector yielded 350 colonies, while double selection of transfected EPC cells with G418 and gancyclovir (Gc) resulted in nearly complete cell death, demonstrating that the PNS procedure is effective in fish cells. Homologous recombination vectors consist of the Xiphophorus melanoma receptor kinase (X mrk(Y)) gene as homologous sequence in addition to the neo and tk genes. Conditions for homologous recombination vector transfection and drug selection were established. After verification of the feasibility of expression of homologous recombination vectors in EPC cells, the first gene targeting experiments were attempted in the Xiphophorus melanoma cell line, PSM. Positive-negative selection of the targeting vector-transfectants led to a low enrichment in this particular cell line. The reasons for the low enrichment in PSM cells were discussed. (C) 2002 Elsevier Science B.V. All rights reserved.

Selection and ultrastructural observation of a high-CO2-requiring mutant of cyanobacterium Synechococcus sp PCC7942

A high-CO2-requiring mutant of Synechococcus sp. PCC7942 las been isolated after chemical mutagenesis of ethyl methane sulphonate (EMS). It was able to grow at 4% CO2, but not under ambient CO2. The initial screening of the mutant showed that the genetic reversion rate was about 10(-7) and death occurred 2 -3 days after being transferred from 4% CO2 to the ambient air. Its photosynthetic dependence on external dissolved inorganic carbon was higher than that of the wild type cells, but its carbonic anhydrase activity was comparatively low. In the ultrastructural level, various types of aberrant carboxysomes appeared in the mutant cells: rod-shaped carboxysomes, irregular carboxysomes and the "empty-inclusion carboxysomes" with increasing number of glycogen granules surrounding the thylakoids. All these alterations indicated that the mutant was defective in utilizing the external CO2. The induction of carboxysomes by lower levels of CO2 and the biogenesis of carboxysomes are herein discussed.

Determining of the delay time for a heating ventilating and air-conditioning plant using multi-weights neurons approach

This paper presents an multi weights neurons approach to determine the delay time for a Heating ventilating and air-conditioning (HVAC) plan to respond to control actions. The multi weights neurons is a fully connected four-layer network. An acceleration technique was used to improve the general delta rule for the learning process. Experimental data for heating and cooling modes were used with both the multi weights neurons and a traditional mathematical method to determine the delay time. The results show that multi weights neurons can be used effectively determining the delay time for HVAC systems.

tilt menu: using the 3d orientation information of pen devices to extend the selection capability of pen-based user interfaces

We present a new technique called‘Tilt Menu’ for better extending selection capabilities of pen-based interfaces.The Tilt Menu is implemented by using 3D orientation information of pen devices while performing selection tasks.The Tilt Menu has the potential to aid traditional onehanded techniques as it simultaneously generates the secondary input (e.g., a command or parameter selection) while drawing/interacting with a pen tip without having to use the second hand or another device. We conduct two experiments to explore the performance of the Tilt Menu. In the first experiment, we analyze the effect of parameters of the Tilt Menu, such as the menu size and orientation of the item, on its usability. Results of the first experiment suggest some design guidelines for the Tilt Menu. In the second experiment, the Tilt Menu is compared to two types of techniques while performing connect-the-dot tasks using freeform drawing mechanism. Results of the second experiment show that the Tilt Menu perform better in comparison to the Tool Palette, and is as good as the Toolglass.

IMPLANTATION ENERGY SELECTION IN MOLECULAR-BEAM EPITAXY GAAS FILMS ON SI SUBSTRATES

GaAs films made by molecular beam epitaxy with thicknesses ranging from 0.9 to 1.25-mu-m on Si have been implanted with Si ions at 1.2 MeV to dose of 1 x 10(15)/cm2. A rapid infrared thermal annealing and white light annealing were then used for recrystallization. Crystalline quality was analysed by using backscattering channeling technique with Li ion beam of 4.2 MeV. The experimental results show that energy selection is important for obtaining better and uniform recrystallized GaAs epilayers.

Spatial Mode Selection by the Phase Modulation of Subwavelength Plasmonic Grating

The extraordinary transmission of the subwavelength gold grating has been investigated by the rigorous coupled-wave analysis and verified by the metal-insulator-metal plasmonic waveguide method. The physical mechanisms of the extraordinary transmission are characterized as the excitation of the surface plasmon polariton modes. The subwavelength grating integrated with the distributed Bragg reflector is proposed to modulate the phase to realize spatial mode selection, which is prospected to be applied for transverse mode selection in the vertical cavity surface-emitting laser.

重离子诱变技术选育碱性蛋白酶高产菌株；The Selection for High-yield Alkaline Protease Producing Strain by Heavy-ion Irradiation

从采集的土壤样品中分离筛选出一株碱性蛋白酶产生菌G-41,经16S rRNA分子鉴定为芽孢杆菌属菌株。该菌株在发酵培养基中能产生较高产量的胞外碱性蛋白酶(1.7×104U/mL)。以G-41为出发菌株,对其进行重离子辐照诱变处理,获得突变株G-41-68,将该突变株再次经重离子诱变,从大量突变株中筛选出碱性蛋白酶高产菌株15Gy-54,其酶活力达到6.22×104U/mL。与出发菌株相比较,突变株G-41-68和15Gy-54的酶活力分别提高了1.58倍和2.65倍。对突变株15Gy-54的发酵条件进行了优化研究,结果表明,该菌株的碱性蛋白酶活力得到进一步提高,达到7.18×104U/mL,其最适发酵条件为:培养基(g/100mL)为胰蛋白胨1、酵母膏0.5、乳糖5、Na2HPO4·12H2O0.4、KH2PO40.03、Na2CO30.1、MgSO40.0481(4×10-3mol/L)、pH8.0,培养温度41℃,振荡培养时间42-48h。实验结果表明,重离子辐照诱变技术是一种非常有效的微生物诱变育种新技术。

基于克隆选择算法的非线性优化迭代学习控制；Clonal selection algorithm based nonlinear optimal iterative learning control

为实现对模型不确定的有约束非线性系统在特定时间域上输出轨迹的有效跟踪,将改进的克隆选择算法用于求解迭代学习控制中的优化问题。提出基于克隆选择算法的非线性优化迭代学习控制。在每次迭代运算后,一个克隆选择算法用于求解下次迭代运算中的最优输入,另一个克隆选择算法用于修正系统参考模型。仿真结果表明,该方法比GA-ILC具有更快的收敛速度,能够有效处理输入上的约束以及模型不确定问题,通过少数几次迭代学习就能取得满意的跟踪效果。