密码子优化的牛精蛋白基因在大肠杆菌中的表达

以PCR的方法得到牛精蛋白基因的基因，去其内含子，得到牛精蛋白cDNA，克隆入原核表达载体pGEX-2T中，组装成表达载体pGEX-2T-PE，利用大肠杆菌偏好的编码精氨酸的密码子CGT将野生型牛精蛋白基因中编码精氨酸的稀有密码子（AGA或AGG）替换掉，通过基因合成得到密码子优化的牛精蛋白的基因，将其克隆入原核表达载体pGEX-2T中，组装成表达载体pGEX-2T-PS。将这两个表达载体分别转化入大肠杆菌表达菌株BL21中，经IPTG诱导，同样条件下，野生型牛精蛋白基因无法得到表达，密码子优化的牛精蛋白基因能够得到良好的表达，表达产物约占细菌总蛋白的18％，将表达蛋白纯化，进行DNA－蛋白结合实验，发现其能与DNA发生非特异性的结合。     

PRO/E在波浪圈成型模制造中的应用

介绍了波浪圈成型模的三维造型与数控加工自动编程的过程 ,并结合实例对PRO E软件的特点进行了分析 ,指出CAD CAM的运用可提高模具加工质量 ,缩短设计制造的周期。     

LXT384接口芯片的几种环回形式

LXT384是一个用于SONET/SDH设备的八进制T1/E1/J1线路接口单元芯片。文中简述了该芯片在实际应用中进行自检的若干种不同的环回形式(包括：模拟环回、数字环回、运程环回等)，直观地指出了各种环回形式的特点和区别。     

采用5V单电源供电的通信收发器LMS202E/LMS485及其应用

介绍了采用单5V电源工作的RS-232通信双收发器LMS202E和高速双向数据通信差分总线收发器LMS485的主要特性 ,给出了它们的典型应用电路。     

Channelling studies on the lattice location of B atoms in graphite

In order to obtain information on the lattice location of B atoms in graphite, channelling experiments have been performed at room temperature with a proton beam of an energy of 0.65-0.77 MeV for the 〈0 0 0 1〉 axial channel in highly oriented pyrolytic graphite (HOPG) crystals doped with 0.32 at.% B. The B atoms are detected by measuring α-particles which are emitted as a result of a nuclear reaction ¹¹B(p,α)αα. It is clearly demonstrated that most of B atoms are shadowed behind the 〈0 0 0 1〉 C atomic rows. Taking account of the already reported experimental results on a change of lattice parameters by B-doping, it is concluded that most of B atoms are located at substitutional sites. It is also observed that B-doping introduces lattice strain on the c-plane. In addition, the presence of a small portion of interstitial B atoms is suggested.     

基于PRO/E三维管道轨迹专用检具的设计

介绍使用该软件组件设计模块中的高级工具———切除 (CutOut)功能设计高精度三维轨迹管道零件检具的方法 ,该方法具有高效和精密等特点。     

Purification and renaturation of recombinant human lymphotoxin (tumour necrosis factor beta) expressed in Escherichia coli as inclusion bodies

High level expression of recombinant human tumour necrosis factor β (rh TNF-β) in Escherichia coli results in the formation of two portions of protein, namely soluble active protein and insoluble protein which is inactive and aggregates in the form of inclusion bodies (IBs). In this study, a procedure for purification and renaturation of rh TNF-β from inclusion bodies has been designed and verified experimentally with a product purity of more than 90% and a recovery of about 30%. The procedure includes washing of IBs with specific wash buffer (Triton X-100/EDTA/lysozyme/PMSF), their solubilization with 8 mol dm^?3 alkaline urea, purification with ion-exchange columns, refolding with renaturation buffer and finally concentration and desalination with an ultrafiltration membrane. The characteristics of the renatured protein were identical with those of purified protein from the soluble fraction as demonstrated by (1) SDS-PAGE, (2) cytotoxic activity on mouse L929 cells, (3) N-terminal amino acid sequence, and (4) gel filtration chromatography.     

An integrated downstream processing strategy for the recovery and partial purification of penicillin acylase from crude media

An integrated process strategy for the recovery of penicillin acylase was developed, based on precipitation of non‐enzymatic proteins directly from Escherichia coli homogenates or crude extracts using Rolquat (quaternary ammonium salt) and adsorption of the enzyme on Amp‐Seph (3.8 µmole ampicillin cm^?3) under pseudo‐affinity conditions. The effect of pH, concentrations of ammonium sulfate and Rolquat, and also concentrations of protein and cell debris on the precipitation of non‐enzymatic proteins from homogenates and crude extracts of penicillin acylase were analysed. The method of addition of Rolquat to homogenates and crude extracts significantly influenced the size of the precipitated particles. Improved results on the specific activity of penicillin acylase were obtained for 22% and 1% (w/v) of ammonium sulfate and Rolquat, respectively, added sequentially to enzyme solutions and at room temperature. Under these experimental conditions, the specific activity of penicillin acylase in homogenates and crude extracts was enhanced 2.5–3.0‐fold. Finally, the integrated process strategy was implemented first by precipitation of non‐enzymatic proteins and recovery of penicillin acylase directly from the enzyme solution treated with Rolquat using an adsorption/filtration system with an overall yield of 86%. This system allows simultaneously the filtration of cell debris and fine precipitated particles, in situ recovery of penicillin acylase by its adsorption on Amp‐Seph, and selective desorption of the enzyme with a specific activity of 11 IU (mg prot)^?1 and a desorption yield of 95%. © 2002 Society of Chemical Industry     

Kinetic investigation of the carbothermal reduction of an Iranian clay

The kinetics of the carbothermal reduction of clay under argon atmosphere has been investigated by the X-ray diffraction method. The clay-carbon (excess) mixture was formed into 2 cm diameter disks of different thicknesses. Experimental data evidences the significant effect of sample thickness on the reaction rate at 1400 °C. Decreasing thickness promotes mullite dissociation and formation of SiC and alumina powders. Mathematical modeling of the reaction system showed the gas diffusion in the Knudsen regime through the pellet to be the rate controlling step. Diffusivity of CO and reacted core tortuosity factor have been calculated.     

Forty-Five Years After Broadbent (1958): Still No Identification Without Attention.

According to D. E. Broadbent's (1958) selective filter theory, people do not process unattended stimuli beyond the analysis of basic physical properties. This theory was later rejected on the basis of numerous findings that people identify irrelevant (and supposedly unattended) stimuli. A careful review of this evidence, however, reveals strong reasons to doubt that these irrelevant stimuli were in fact unattended. This review exposed a clear need for new experiments with tight control over the locus of attention. The authors present 5 such experiments using a priming paradigm. When steps were taken to ensure that irrelevant stimuli were not attended, these stimuli produced no priming effects. Hence, the authors found no evidence that unattended stimuli can be identified. The results support a modern version of Broadbent's selective theory, updated to reflect recent research advances. (PsycINFO Database Record (c) 2010 APA, all rights reserved)