苔藓植物DNA提取方法研究

提取高质量的 DNA是对苔藓植物遗传多样性进行研究的基础。该文以苔藓植物为试材 ,用 5种方法 ,即快速提取法、改良 CTAB法、CTAB法、SDS法及高盐法 (第一种为自行设计 ,第二种是对原有方法的改进 )对苔藓植物 DNA提取方法进行了比较研究。结果表明 ,快速提取法和改良 CTAB法是 2种适合于苔藓植物 DNA提取的方法。这 2种方法提取的 DNA浓度和纯度均比较高 ,凝胶电泳显示无明显降解现象 ,适宜作为 PCR扩增的模板 ,并成功地进行了 RAPD扩增。

提取北方土壤真菌DNA的一种方法

由于土壤理化性质的复杂性和真菌细胞壁结构的特殊性,从土壤样品中提取真菌基因组DNA比较困难。中国北方土壤与其它地区土壤相比有其自身的特点,因此,有必要优化一种适合于北方土壤真菌DNA提取的方法。本实验向灭菌黑土中分别投加12种在系统分类上差别较大的真菌,以传统土壤总DNA提取方法及纯菌DNA提取方法为基础,分别与蜗牛酶,纤维素酶进行组合、优化,得到7种不同的土壤真菌基因组DNA提取方法。利用真菌28SrDNA通用引物U1/U2-GCPCR-DGGE分析方法分别考察了7种不同方法所提取土壤真菌基因组DNA的多样性和代表性。结果表明:1)液氮研磨,纤维素酶、蜗牛酶和溶菌酶(浓度分别为6、3和1mg.ml-1)37℃作用60min,2%SDS于65℃裂解30min;2)-65℃～65℃冻融3次,纤维素酶、蜗牛酶和溶菌酶(浓度分别为6、3和1mg.ml-1)37℃作用180min,2%SDS于65℃裂解30min的组合具有较好的提取效果。利用后一种方法分别对3种理化性质差异较大的中国北方自然土壤样品真菌DNA进行提取并分析,表明所提取土壤基因组DNA真菌特异性PCR-DGGE图谱条带丰富,该方法可用于多种北方土壤真菌多样性研究。

鸭瘟病毒gB蛋白N端抗原域的高效表达及其免疫特性

在分析鸭瘟病毒(Duck plague virus,DPV)gB蛋白抗原性的基础上,设计1对引物克隆gB蛋白N端抗原性较好的抗原域编码基因,克隆到表达载体pET32a中,构建了原核表达质粒pET-gB1。将pET-gB1转化到感受态大肠杆菌(Escherichia coli)BL21(DE3)中,经IPTG诱导和SDS-PAGE分析,可见约42.4kD的目的蛋白以包涵体形式表达。Western blot分析发现,表达产物与抗鸭瘟的鼠阳性血清发生特异性反应。将包涵体溶解于8mol/L的尿素中,利用His·Bind试剂盒获得纯化的蛋白,将纯化的蛋白皮下注射免疫小鼠,间接ELISA法测得抗体的效价,MTT法检测免疫小鼠的T淋巴细胞增殖反应能力。结果说明,该融合蛋白能够诱导机体产生较强的体液免疫和细胞免疫。

reuse strategies in distributed complex event detection

Software Engineering Society of Korean; Institute for Information Scientists and Engineers; IEEE Reliability Society; KAIST (Korea Advanced Institute of Science and Technology); Korea Information Promotion Agency; Samsung SDS

Low Cationic Proportion Ampholytic Polymer: Synthesis,Solution Properties and Interaction with Anionic Surfactant

Novel ampholytic terpolymer of N-vinylformamide (NVF), vinylamine (VAm) and sodium acrylate (NA) with low cationic proportion was obtained by hydrolyzing copolymer of NVF and NA (PNVFNA). Solution properties of the polymer were investigated by methods of turbidity and viscosity experiment. The effect of sodium dodecyl sulfate (SDS) on solution viscosity was also investigated. The results showed that the turbidity curves were bimodal, and pH 3.0 was determined as the isoelectric point (IEP).

Sensitive, Label-Free Protein Assay Using 1-ethyl-3-methylimidazolium Tetrafluoroborate Supported Microchip Electrophoresis with Laser-Induced Fluorescence Detection

Based on the dimer-monomer equilibrium movement of the fluorescent dye Pyronin Y (PY), a rapid, simple, highly sensitive, label-free method for protein detection was developed by microchip electrophoresis with LIF detection. PY formed a nonfluorescent dimer induced by the premicellar aggregation of an anionic surfactant, SDS, however, the fluorescence intensity of the system increased dramatically when proteins such as BSA, bovine hemoglobin, cytochrome c, and trypsin were added to the solution due to the transition of dimer to fluorescent monomer. Furthermore, 1-ethyl-3-methylimidazolium tetrafluoroborate (EMImBF(4)) instead of PBS was applied as running buffers in microchip electrophoresis.

Separation and detection of narcotic drugs on a microchip using micellar electrokinetic chromatography and electrochemiluminescence

A new approach for fast and sensitive electrochemiluminescence (ECL) detection of narcotic drugs on a microchip after separation by micellar electrokinetic chromatography (MEKC) is presented, taking the cocaine and its hydrolysate ecgonine as the test analytes. The mixture of hydrophilic BMIMBF4 ionic liquid (IL) and sodium dodecyl sulfate (SDS) was used directly as the buffer of MEKC with less noisy baselines, lower electrophoretic current and satisfactory separation performance.

Self-assembled 3D microflowery In(OH)(3) architecture and its conversion to In2O3

Using sodium dodecyl sulfate (SDS), a 3D microflowery indium hydroxide [In(OH)(3)] structure assembled from 2D nanoflakes was fabricated in a large quantity via a hydrothermal approach at relative low temperature. The obtained In(OH)(3) flowers exhibited a narrow size range between 4 and 6 mu m. The properties of these composites were characterized by XRD, EDX, FE-SEM, TEM, SAED, and TGA. In this work, both the use of urea and SDS and the amounts of these components played important roles in the formation of In(OH)3 with different nanostructures.

Synthesis of mesoporous LaPO4 nanostructures with controllable morphologies

Lanthanum phosphate (LaPO4) nanostructures with different morphologies were prepared by a facile solution-precipitation process. The effect of different reaction conditions on the morphology of nanostructures was studied. When the molar ratio of La3+:H3PO4 was around 1 : 2, 1 : 20, 1 : 100, and 1 : 200, four different morphologies, such as near-spherical, snowflake-like, star-shaped, lens-like nanostructures and short nanorods, were obtained, respectively. Meanwhile, similar shapes developed when the molar ratio of H3PO4 to ionic surfactants, such as SDS and CTAB, was varied. In addition, Eu3+ doped and Ce3+/Tb3+ co-doped LaPO4 nanostructures showed morphology evolution similar to undoped LaPO4 nanostructures. The optical properties of these doped LaPO4 were also characterized.

中药羚羊角及其类似品中水溶性蛋白的检测

目的:研究中药羚羊角及其类似品中水溶性蛋白组分差异及鉴定方法,为其药效组分质量评价体系的建立提供科学数据。方法:采用紫外分光光度法(UV)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,对羚羊角及其类似品山羊角和黄羊角进行水溶性蛋白质含量和组分对比分析。结果:羚羊角中水溶性蛋白的含量高于黄羊角和山羊角,且3种角类中药蛋白组分的组成差异显著。结论:蛋白组分可用于羚羊角的鉴定,采用药效蛋白组分评价角类中药质量具有明显的科学性和实用性。

Purification, sequencing and biological activity of polypeptide from velvet antler

The velvet antler polypeptide CNT14 was extracted and purified by gel filtration, ion exchange chromatography and RP C, which showed a single peak in HPLC chromatography and a single band in SDS-PAGE. The molecular weight measured by MALDI/TOF/MS spectrum was 1479. 9028. The polypeptide consisted mostly of Glu, Leu, Val, Pro. The amino acid sequence of the polypeptide was detected with ESI-MS/ MS, and the sequence was E-P-T-V-L-D-E-V-C-L-A-H-G-P. The experiments of biological activity of polypeptide CNT14 in vivo were carried out, and the results show that CNT14 has stimulant effects on the growth of rat HT22 cells. Then we produced the polypeptide CNT14 according the amino acid sequence by solid phase synthesis, confirmed the sequence of the polypeptide to be consistent with the amino acid sequence of polypeptide CNT14 which was separated from the velvet antler.

在线纯化技术应用于MALDI-TOFMS测定溶菌酶的分子量

在用基质辅助激光解吸电离飞行时间质谱(MALDI-TOFMS)测定蛋白质分子量的过程中,一些盐和蛋白质变性剂经常大大抑制样品信号,产生一些难以解析的离子峰,因此测试前应尽可能去除样品中的添加剂。为此,本研究建立了MALDI—TOFMS测试中在线纯化蛋白质样品的新方法。采用硝酸纤维素膜作为固相载体,将标准蛋白质溶菌酶制成含6 mol/L盐酸胍变性剂、2％SDS表面活性剂的100 mmol/L Tris—HCL溶液进行质谱测定。结果表明:新方法简单、快速,可明显增强离子峰的强度,提高测定蛋白质分子量的灵敏度。

Enzymatic degradation of poly(L-lactide) and poly (epsilon-caprolactone) electrospun fibers

Poly(L-lactide) (PLLA) and poly(epsilon-caprolactone) (PCL) ultrafine fibers were prepared by electrospinning. The influence of cationic and anionic surfactants on their enzymatic degradation behavior was investigated by measuring weight loss, molecular weight, crystallinity, and melting temperature of the fibers as a function of degradation time. Under the catalysis of proteinase K, the PLLA fibers containing the anionic surfactant sodium docecyl sulfate (SDS) exhibited a faster degradation rate than those containing cationic surfactant triethylbenzylammonium chloride (TEBAC), indicating that surface electric charge on the fibers is a critical factor for an enzymatic degradation. Similarly, TEBAC-containing PCL fibers exhibited a 47% weight loss within 8.5 h whereas SDS-containing PCL fibers showed little degradation in the presence of lipase PS. By analyzing the charge status of proteinase K and lipase PS under the experimental conditions, the importance of the surface charges of the fibers and their interactions with the charges on the enzymes were revealed. Consequently, a "two-step" degradation mechanism was proposed: (1) the enzyme approaches the fiber surface; (2) the enzyme initiates hydrolysis of the polymer.

Preparation and properties of abzyme with thyroxine deiodinase

The abzyme (Se-6E8) with a higher thyroxine deiodinase activity was prepared by modifying the serine residues of monoclonal antibody (6E8)with phenylmethanesulfonyl fluoride and sodium hydrogen selenide, and the 6E8 against O-methyl-T-4, which is a kind of thyroxine derivatives and was taken as a hapten for the first time. Two bands were found corresponding to the 5.5 kD heavy chain and the 2.7 kD light chain respectively by SDS-PAGE. The characteristics of dissociation constants, pH, and temperature were also studied. The results show that the activity of Se-6E8 is 2 010 U/mumol protein, and the proper temperature and pH of the catalytic reactions is 57 degreesC and 8.2 respectively.

High-sensitivity molecular mass determination of lysozyme prior to MALDI-MS

Sodium dodecyl sulfate(SDS) is a powerful solubilizing detergent which is often used during the separation of highly complex protein mixtures by one- or two-dimensional (2D) gel electrophoresis. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a widely used technique for mass spectrometric analysis of some protein molecules compared to other techniques. But the presence of SDS or some salts usually leads to signal deterioration when using MALDI-MS. A method for using nitrocellulose membrane as the solid-phase carrier combined with n-octyl-beta-D-glucopyranoside in the matrix highly enhances the sensitivity of the molecular mass determination of lysozyme. This technique has the advantage that the signal-to-noise of the molecular weight profile is improved compared with the mass spectrum and the profile is relatively easy to interpret.