Trypsiligase-Catalyzed Labeling of Proteins on Living Cells

Fluorescent fusion proteins are powerful tools for studying biological processes in living cells, but universal application is limited due to the voluminous size of those tags, which might have an impact on the folding, localization or even the biological function of the target protein. The designed biocatalyst trypsiligase enables site-directed linkage of small-sized fluorescence dyes on the N terminus of integral target proteins located in the outer membrane of living cells through a stable native peptide bond. The function of the approach was tested by using the examples of covalent derivatization of the transmembrane proteins CD147 as well as the EGF receptor, both presented on human HeLa cells. Specific trypsiligase recognition of the site of linkage was mediated by the dipeptide sequence Arg-His added to the proteins’ native N termini, pointing outside the cell membrane. The labeling procedure takes only about 5 minutes, as demonstrated for couplings of the fluorescence dye tetramethyl rhodamine and the affinity label biotin as well.     

Fabrication and joining mechanism of Nano-Cu/Si3N4 ceramic substrates

A novel method for fabricating a nano-Cu/Si₃N₄ ceramic substrate is proposed. The nano-Cu/Si₃N₄ ceramic substrate is first fabricated using spark plasma sintering (SPS) with the addition of nanoscale multilayer films (Ti/TiN/Ti/TiN/Ti) as transition layers. The microstructures of the nano-Cu metal layer and the interface between Cu and Si₃N₄ are investigated. The results show that a higher SPS temperature increases the grain size of the nano-Cu metal layer and affects the hardness. The microstructure of the transition layer evolves significantly after SPS. Ti in the transition layer can react with Si₃N₄ and with nano-Cu to form interfacial reaction layers of TiN and Ti–Cu, respectively; these ensure stronger bonding between nano-Cu and Si₃N₄. Higher SPS temperatures improve the diffusion ability of Ti and Cu, inducing the formation of Ti₃Cu₃O compounds in the nano-Cu metal layer and Ti₂Cu in the transition layer. This study provides an important strategy for designing and constructing a new type of ceramic substrate.     

Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

二氧化锡溅射工艺研究

SnO2 是最早使用也是目前使用最广泛的一种气敏材料 ,使用该材料设计制作的气敏传感器具有许多优点。在简要介绍溅射镀膜的成膜过程和特点的基础上 ,着重介绍了SnO2 膜的制备流程 ,分析了功率和温度变化对成膜质量的影响     

彩色PDP用玻璃基板

本文讨论了彩色ＰＤＰ现用钠钙玻璃的热稳定性，指出由于这种玻璃在热处理过程中易变形和收缩，因而不适合作大面积彩色ＰＤＰ基板材料，介绍了彩色ＰＤＰ基板玻璃的制造方法和特点。最后介绍了日本旭硝子公司和美国康宁公司各自为彩色ＰＤＰ新开发的玻璃基板材料ＰＤ２００和ＣＳ２５。     

晶体硅薄膜电池制备技术及研究现状

晶体硅薄膜太阳电池近些年来得到广泛的研究和初步的商业化探索。根据所采用的晶体硅薄膜沉积工艺中温度范围的不同，晶体硅薄膜电池研究可分为高温路线和低温路线两个不同发展方向。本文分别从这两个方向综述了目前国外晶体硅薄膜电池制备技术的最新进展，最新实验室研究结果。报导了晶体硅薄膜电池商业化进展状况，指出了晶体硅薄膜电池实现产业化必须解决的问题。     

针刺合成革基布质量的探讨

分析了针刺合成革基布质量的控制要素 ,以及预针刺机的牵伸比和针刺机的步进图对基布质量的影响     

可与氟树脂基板相匹敌的热可塑性树脂基板材料

概述了可以与氟树脂基板相匹敌的低介质特性的热可塑性树脂基板材料的开发和应用。     

流延法制作AlN陶瓷基片工艺

研究出一种用流延法制作ＡＩＮ陶瓷基片的工艺。制备出性能优良的ＡＩＮ基片。经１６５０℃，保温２ｈ无压烧结制得的基片，其热导率达１３０Ｗ／（ｍ·Ｋ）、密度为３．３４ｇ／ｃｍ￣３，表面光滑平整，瓷体结构均匀，性能达到用干压法制备的同类基片的性能。     

Kinetic analysis for effect of initial substrate concentration on growth and secondary metabolite production in cultures of Nicotiana tabacum

Suspension cultures of tobacco cells were studied using airlift and rotary-drum bioreactors. The effect of initial concentrations of a major substrate, sucrose, on the growth and production of a secondary metabolite, phenolic compounds, was investigated. The dry weights and total concentrations of the phenolic compounds increased with the initial sucrose concentration in both bioreactors. Both bioreactors were found to have the same tendency for the effect of initial sucrose concentration. The structured model, presented previously was modified by considering that sucrose was hydrolyzed to glucose and fructose by an enzymatic reaction. The previous and the new models were applied to the above two sets of experimental data obtained with two bioreactors, independently. The hydrolysis of sucrose was elucidated to contribute slightly to the overall kinetics of growth and secondary metabolite production in these cultures. Furthermore, the levels of shear damage in each bioreactor were quantitatively compared based on the death rate constant, k_i, which is one of the model constants.