De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase

Classical swine fever virus (CSFV) non-structural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp), a key enzyme which initiates RNA replication by a de novo mechanism without a primer and is a potential target for anti-virus therapy. We expressed the NS5B protein in Escherichia coli. The rGTP can stimulate de novo initiation of RNA synthesis and mutation of the GDD motif to Gly-Asp-Asp (GAA) abolishes the RNA synthesis. To better understand the mechanism of viral RNA synthesis in CSFV, a three-dimensional model was built by homology modeling based on the alignment with several virus RdRps. The model contains 605 residues folded in the characteristic fingers, palm and thumb domains. The fingers domain contains an N-terminal region that plays an important role in conformational change. We propose that the experimentally observed promotion of polymerase efficiency by rGTP is probably due to the conformational changes of the polymerase caused by binding the rGTP. Mutation of the GDD to GAA interferes with the interaction between the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive. (c) 2005 Elsevier B.V. All rights reserved.

Dewetting of polymethyl methacrylate on the patterned elastomer substrate by solvent vapor treatment

The dewetting evolution process of polymethyl methacrylate (PMMA) film on the flat and prepatterned polydimethylsiloxane (PDMS) substrates (with square microwells) by the saturated solvent of methyl ethyl ketone (MEK) treatment has been investigated at room temperature by the optical microscope (OM) and atomic force microscope (AFM). The final dewetting on the flat PDMS substrate led to polygonal liquid droplets, similar to that by temperature annealing. However, on the patterned PDMS substrate, depending on the microwells' structure of PDMS substrate and defect positions that initiated the rupture and dewetting of PMMA, two different kinds of dewetting phenomena, one initiated around the edge of the microwells and another initiated outside the microwells, were observed. The forming mechanism of these two different dewetting phenomena has been discussed. The microwells were filled with liquid droplets of PMMA after dewetting due to the formation of fingers caused by the pinning of the three-phase-line at the edge of the microwells and their rupture.

Identification and expression of TRAF6 (TNF receptor-associated factor 6) gene in Zhikong Scallop Chlamys farreri

Tumor necrosis factor receptor-associated factor 6 (TRAF6), a key signaling adaptor molecule common to the TNFR superfamily and IL-IR/TLR family, is important not only for a diverse array of physiological processes functions of the TNFR superfamily, but also is involved in adaptive immunity and innate immunity. In this report, the first bivalve TRAF6 (named as CfTRAF6) gene is identified and characterized from Zhikong scallop Chlamys farreri. The full-length cDNA of CfTRAF6 is of 2510 bp, consisting of a 5'-terminal untranslated region (UTR) of 337 bp, a 3'-terminal UTR of 208 bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame (ORF) encoding a polypeptide of 655 amino acids. The predicted amino acid sequence of CfTRAF6 comprises characteristic motifs of the TRAF proteins, including a Zinc finger of RING-type, two Zinc fingers of TRAF-type, a coiled-coil region, and a MATH (the meprin and TRAF homology) domain. The overall amino acid sequence identity between CfTRAF6 and other TRAF6s is 28-68%. Phylogenetic analyses of CfTRAF6 sequence with TRAF sequences from other organisms indicate that CfTRAF6 is a true TRAF6 orthologue. The mRNA expression of CfTRAF6 in various tissues is measured by Real-time RT-PCR. The mRNA transcripts are constitutively expressed in tissues of haemocyte, muscle, mantle, heart, gonad and gill, but the highest expression is observed in the gonad. The temporal expressions of CfTRAF6 mRNA in the mixed primary cultured haemocytes are recorded after treatment with 20 mu g mL(-1) and 0.5 mu g mL(-1) peptido-glycan (PGN). The expression level of CfTRAF mRNA is down-regulated from 1.5 h to 3 h after the treatment with 0.5 mu g mL(-1) PGN, and then recovers to the original level. While the expression of CfTRAF6 is obviously decreased after treatment with 20 mu g mL(-1) PGN, and reach the lowest point (only about 1/9 times to control) at 3 h. The result Suggests that CfTRAF6 can be greatly regulated by PGN and it may be involved in signal transduction and immune response of scallop. (C) 2008 Published by Elsevier Ltd.

用光学阵列传感器的机器人物体分类系统

本文介绍用光学阵列传感器的机器人物体分类系统。传感器直接安装在机器人的两个手指上。被抓物体的阴影通过光导纤维传到安放在“安全区”的光敏元件上。计算机识别物体的轮廓后命令机器人抓握物体,并把它运送到指定的地点从而达到物体分类的目的。

构造复杂区变速成图方法研究

In the complex structure areas, velocity field building and structure mapping are important for seismic exploration. With the development of seismic exploration, the methods of structure mapping, reservoir prediction and reservoir description all require high precious velocity field. And more accurate depth-structure maps are required for well site design. Aiming at the problems and defects in velocity analysis and structure mapping in oil seismic exploration, the paper which is based on the studies of real data in several areas combines the theories with practical application, and analyzes the precision and applicability of several methods of velocity model building. After that, the following methods are mainly studied: the coherence inversion methods based on the pre-stack CMP gathers or stacking velocity; the interval velocity inversion methods constrained by multi-well; the Random Simulation method; 3D Image Ray Map Migration method and the structure mapping in floating datum and in fixed datum, and then we conclude the method of building high precious seismic velocity field and structure mapping with variable velocity. Firstly, the paper analyses the distributing rule of the velocity variation in the areas with complex structures in the northwest of China, then points out that velocity is a crucial factor which influences the precision of structure mapping, and the velocity variations have something to do with the shapes of the structures, the variety of lithology and so on. The key point of improving the precision of seismic velocity field is to obtain a structure mapping with high precision. We also describe the range and conditions of these methods. Secondly, by comparing many popular methods of velocity model building, we propose a new method in the use of velocity model building. The new method is more effective in velocity model building under every kind of complex condition and is worthy of spreading. At last, the paper fingers out that it is a system engineering to study variable velocity mapping in every kind of complex structure areas. Every step of the work can affect the final results. So it is important to build high efficient and practical velocity model and the flows of mapping processing. The paper builds the flows and gives some examples. The method has been applied in more than ten exploring surveys. The application proves that this method could bring good effect on researching on low-amplitude trap, reservoir prediction, reservoir description and the integrated research of oil&gas geology. Keywords: structure mapping velocity model building complex structure variable velocity media