Integration of double-fluorescence expression vectors into zebrafish genome for the selection of site-directed knockout/knockin

Production of zebrafish by modifying endogenous growth hormone (GH) gene through homologous recombination is described here. We first constructed the targeting vectors pGHT1.7k and pGHT2.8k, which were used for the knockout/knockin of the endogenous GH gene of zebrafish, and injected these two vectors into the embryos of zebrafish. Overall, the rate of targeted integration with the characteristic of germ line transmission in zebrafish was 1.7x10(-6). In one experimental patch, the integrating efficiency of pGHT2.8k was higher than that of pGHT1.7k, but the lethal effect of pGHT2.8k was stronger than that of pGHT1.7k. The clones with the correct integration of target genes were identified by a simple screening procedure based on green fluorescent protein (GFP) and RFP dual selection, which corresponded to homologous recombination and random insertion, respectively. The potential homologous recombination zebrafish was further bred to produce a heterozygous F-1 generation, selected based on the presence of GFP. The potential targeted integration of exogenous GH genes into a zebrafish genome at the P-0 generation was further verified by polymerase chain reaction and Southern blot analysis. Approximately 2.5% of potential founder knockout and knockin zebrafish had the characteristic of germ line transmission. In this study, we developed an efficient method for producing the targeted gene modification in zebrafish for future studies on genetic modifications and gene functions using this model organism.

Materials selection in mechanical design for microsensors and microactuators

Microsensors and microactuators are vital organs of microelectromechanical systems (MEMS), forming the interfaces between controller and environment. They are usually used for devices ranging in size at sub-millimeter or micrometer level, transforming energy between two or more domains. Presently, most of the materials used in MEMS devices belong to the silicon material system, which is the basis of the integrated circuit industry. However, new techniques are being explored and developed, and the opportunities for MEMS materials selection are getting broader. The present paper tries to apply 'performance index' to select the material best suited to a given application, in the early stage of MEMS design. The selection is based on matching performance characteristics to the requirements. A series of performance indices are given to allow a wide range comparison of materials for several typical sensing and actuating structures, and a rapid identification of candidates for a given task. (C) 2002 Elsevier Science Ltd. All rights reserved.

Effect of surface tension on the mode selection of vertically excited surface waves in a circular cylindrical vessel

Singular perturbation theory of two-time-scale expansions was developed in inviscid fluids to investigate patternforming, structure of the single surface standing wave, and its evolution with time in a circular cylindrical vessel subject to a vertical oscillation. A nonlinear slowly varying complex amplitude equation, which involves a cubic nonlinear term, an external excitation and the influence of surface tension, was derived from the potential flow equation. Surface tension was introduced by the boundary condition of the free surface in an ideal and incompressible fluid. The results show that when forced frequency is low, the effect of surface tension on the mode selection of surface waves is not important. However, when the forced frequency is high, the surface tension cannot be neglected. This manifests that the function of surface tension is to cause the free surface to return to its equilibrium configuration. In addition, the effect of surface tension seems to make the theoretical results much closer to experimental results.

The mechanism of pattern selection in coupled map lattices

The pattern selection of one-dimensional coupled map lattices is studied in this paper. It is shown by spatiotemporal variable separation that there exists a threshold wavelength in pattern selection which possesses wave-like structures in space and periodic chaotic motion in time.