植物抗病防卫基因及其顺式元件的利用

植物抗病防卫基因的表达产物直接参加对病原物侵染的抵抗活动。这类基因都要由不同刺激信号诱导而表达，基因顺式调控元件尤其是激应性元件（ＲＥ）起关键作用。在ＲＥ可以接收的刺激信号中，化学诱导物是人们操纵基因表达、诱发抗病性的有效工具。根据防卫基因的表达调控机制，病原真菌的致病机制和植物的抗病机，一种在化学诱导型ＲＥ、强启动子和增强子调控下的防卫基因重组体的构建，将在转化作物，改造对真菌病毒的抗性方面发挥     

基因枪法建立紫菜丝状体遗传转化模式

以坛紫菜和条斑紫菜的自由丝状体为材料,利用基因枪法分别转化CaMV35S、SV40、FCP、Amt、Ubi 5种启动子与报告基因组合,转化后48h进行原位组织化学染色检测,首次针对重要经济红藻紫菜的自由丝状体建立并优化遗传转化技术,为紫菜研究提供了新工具。结果显示,SV40启动子可以驱动laeZ报告基因(编码β-半乳糖苷酶)在紫菜丝状体中的瞬间表达,空白与阴性对照未检测到本底;其它4种启动子未检测到基因表达。进一步优化实验发现,在可裂膜650psi、轰击距离6cm下获得最高转化效率为8.0×10~(-5),经定量检测与双因子方差分析,是最佳转化参数,提示基因枪参数对外源基因转化效率具显著影响。     

部分缺失的棉花曲叶病毒互补链基因启动子具有超强活性

棉花曲叶病毒（CLCuV)互补链基因启动子是一种新近鉴定的启动子,它能驱动外源基因在植物体内高效表达,为了研究其最佳启动子区域,对启动子5端进行了一系列缺失,得到5种不同长度的启动子片段与GUS基因融合的植物表达载体,继而导入根癌农杆菌,采用叶盘法转化烟草,并检测转基因植株的GUS活性,实验结果表明,自启动子5端缺失至翻译起始位点上游－287,－271时启动子活性分别是全长启动子的5倍,3倍。首次对棉花曲叶病毒互补链基因启动子的功能区进行了分析比较,发现缺失负调控元件的启动子比全长启动子具有更强的活性,平均活性是CaMV35S启动子的12倍,暗示该启动子具有应用潜力。     

热诱导FLP重组酶删除转基因烟草外源基因

利用热激启动子Hsp18.2、FLP重组酶基因及其专一识别位点构建了重组酶删除系统的植物表达载体转化烟草.热激处理后转基因植株中FLP重组酶表达并识别两个同向loxP-frt重组酶识别位点,使两位点间的DNA插入片段(包含kn1、gus、nptⅡ基因)从转基因烟草基因组中删除,由kn1基因引起的转基因烟草特殊表型及gus基因产生的蓝色表型消失.     

一种新型双向启动子--棉花曲叶病毒启动子的分离、序列分析与功能初探

以棉花曲叶病毒(CLCuV)侵染的烟草叶片组织总DNA为模板,通过聚合酶链反应扩增CLCuV双向启动子片段并插入克隆载体.序列分析和同源性比较表明,克隆的启动子长436bp,与目前发现的9种CLCuV株系的启动子序列均不相同,同源性最高达99.32%.将启动子片段分别以不同方向与GUS报告基因和nos终止子融合,构建了瞬时表达载体.通过基因枪法将质粒载体导入烟草和棉花叶片细胞中进行瞬时表达,结果表明,互补链基因方向启动子属强启动子,在叶肉及维管组织均有较高的活性;病毒链基因方向启动子表达活性较低.本文初步证实分离的互补链基因启动子可作为新型强启动子应用于双子叶植物尤其棉花的遗传转化.     

藻蓝蛋白对Hela细胞CD59基因表达调控作用的研究

探讨了钝顶螺旋藻藻蓝蛋白(PC)对Hela细胞CD59基因表达的调控作用.以正常人CD59cDNA基因为模板,经PCR扩增后重组入真核表达质粒载体pALTER-MAX,然后利用阳离子脂质体(Lipfectamine-2000)将重组质粒和PcDNA共转染人子宫颈癌细胞(Hela)和对照用正常中国仓鼠卵巢细胞(Chinese hamster ovary,CHO)进行表达.用不同浓度的钝顶螺旋藻藻蓝蛋白作用于转染细胞,通过核酸分子杂交技术、免疫荧光标记法和ELISA法对细胞中CD59分子的表达进行检测.结果表明:成功构建了重组质粒pALTER-MAX-CD59,并将其导入真核细胞(Hela,CHO),经G418筛选获得了CD59分子高效表达的细胞克隆.用藻蓝蛋白作用于筛选出的转基因细胞,证实藻蓝蛋白可促进Hela细胞表面CD59蛋白的表达并抑制Hela细胞的增殖,而对于正常CHO细胞无明显作用.     

龙须菜藻红蛋白亚基基因的克隆及其在大肠杆菌中的表达

用PCR扩增方法得到真核红藻－龙须菜藻红蛋白亚基基因序列,与其它6种已知红藻相应序列比较,显示了很高的保守性。在7株藻中β亚基比α亚基保守,α亚基倾向于小区域保守,而β亚基倾向于大片段的保守。该蛋白必需氨基酸的含量较高。藻红蛋白亚基基因在大肠杆菌中达到较高的表达量,诱导4小时,特异表达产物占菌体可溶性蛋白的44％,表达产物呈溶解状态,而两亚基可能是分别翻译的。     

小麦与高冰草体细胞杂种耐盐新品系的盐胁迫应答cDNA差异表达分析

用银染DDRT-PCR技术分析了小麦济南177和小麦与高冰草体细胞杂种耐盐品系杂3号在盐胁迫前后基因的差异表达情况。并通过反向Northern杂交筛选,获得27个阳性片段。从中选出10个候选片段进行序列分析,这些序列信息表明,其表达与盐胁迫应答有一定关系。Northern杂交验证了差异片段wrsi-1在耐盐品系杂3号的根中受盐诱导,可能为胁迫早期应答基因。     

绿色木霉AS3．3711的葡聚糖内切酶V基因的克隆与表达

采用RT-PCR方法克隆到绿色木霉(Trichoderma viride)AS3．3711的葡聚糖内切酶V(EGV)的cDNA基因。测序后构建到酿酒酵母诱导型表达载体pYES2上，转化酿酒酵母，同时研究了超声波处理对酵母完整细胞转化的影响。转化子用2％的β-D-半乳糖进行诱导，用Northem杂交和CMC糖化力法分别对目的基因的转录和表达产物的葡聚糖内切酶活性进行检测。结果表明，EGV的cDNA基因开放阅读框长度为741bp，编码247个氨基酸，推测的蛋白质分子量为24．99kDa。超声波处理60s的酿酒酵母的转化率最高，在相同条件下是未处理组的2倍。酶活检测显示该基因能在酿酒酵母中表达并分泌到胞外。发酵液中的酶活在培养60小时达到最高0．046U／ml。最适酶解温度为60℃，最适pH值为5．4。     

兰花圆球茎基因枪转化后的GUS基因表达

利用花榔菜花叶病毒３５Ｓ启动子引导的葡糖醛酸糖苷酸基因检测基因枪法转化兰花圆球茎组织的瞬间表达效率。发现在基因枪轰击时在ＤＮＡ－金粉悬液中加入０．１ｍｏｌ／Ｌ的生长激素ＮＡＡ，轰击前预培养时使用０．１ｍｏｌ／Ｌ甘露醇处理，每个培养皿轰击压力为１１００ｐｓｉ下，ＧＵＳ基因在兰花圆球茎组织中的瞬间表达效率最高。     

The Research of Anti-Wear Properties of Organomolybdenum Compounds in the Engine Oils

A four-ball tester was used to evaluate the anti-wear performance of three kinds of organomolybdemun compounds in the engine oils, i. e., molybdenum dialkyldithiophosphate (MoDDP), molybdenum dialkyldithiocarbamate ( MoDTC), and sulphur and phosphorus freeorganomolybdeum (Molybdate). The results indicate that a low concentration of MoDDP doesn' t improve the anti-wear properties of the commercial engine oils, but a high concentration of MoDDP can obviously improve the anti-wear properties and the load-carrying capacity of the engine oils. MoDTC doesn' t improve the antiwear properties of the engine oils, but worsens the anti-wear properties of the oils. Signifi can timprove ment of frictional and wear characteristics is obtained with Molybdate added in the commercial engine oils and the formulated oils.     

Certification Becomes Important for the I Quality; and Safety Supervision of the Chinese Liquor Sector

The end of 2007, over 200 unit products of more than 80 Chinese firms have passed the quality grade certification for liquor products. These products involve distilled spirits, beer, wine, yellow wine, fruit syrup wine and others, and cover over 80% of the national top-branded liquor products.……     

Deposition and Coating Properties on CVD Tungsten

Surface characterization and microstructure studies are performed on chemical vapor deposited (CVD) tungsten coating. There is about 2 μm thickness diffusion layer of tungsten in the molybdenum substrate. The thermal shock test shows tungsten coating has good adhesion with molybdenum substrate, but the elements of oxygen and carbon in the tungsten coating have the bad affection to the adhesion. The result of high-temperature diffusion experiment is the diffusion rate from molybdenum substrate to tungsten coating is faster.     

Analysis of marine hydrobiont lipid extracts as possible cryoprotective agents

We studied a combined effect of the cryoprotectants both of lipid and carbohydrate origin at the presence of DMSO or without it on cell viability and the RNA synthesis in the embryonic mollusc and echinoderm cell cultures obtained at different developmental stages. Cryoprotective properties of exogenous lipids correlated with their thermotropic behavior. Lipid extracts from marine hydrobiontes, which thermal transitions are characterized by relatively small part of calorimetric thermogram area lying at temperatures above 0 °C (6–18%), were found to possess a considerable cryoprotective activity. The results are discussed in relation to the prediction of the cryoprotective activity of lipid extracts for marine animal cells. In addition, it was found that marine invertebrate cells can be cryopreserved without DMSO at the presence of trehalose and lipid extracts of some marine hydrobiontes.     

Low-Temperature Properties of Silver

Pure silver is used extensively in the preparation of high-temperature superconductor wires, tapes, films, and other configurations in which the silver not only shields the superconducting material from the surrounding materials, but also provides a degree of flexibility and strain relief, as well as stabilization and low-resistance electrical contact. Silver is relatively expensive, but at this stage of superconductor development, its unique combination of properties seems to offer the only reasonable means of achieving usable lengths of conductor. In this role, the low-temperature physical (electrical, thermal, magnetic, optical) and mechanical properties of the silver all become important. Here we present a collection of properties data extracted from the cryogenic literature and, to the extent possible, selected for reliability.     

Early ironwork in Biscay: Survey,excavation, experimentation and materials characterization. An integral study of the mountainside ironworks (ferrerías de monte or “haizeolak”)

ABSTRACT

The production of iron in Biscay (Basque Country, Spain) has been a hallmark of this territory of the North of the Iberian Peninsula throughout its history. The Archaeology team of the Basque Country Mining Museum has spent 13 years, so far, studying the origins of this activity through the evidences left at the old pre-hydraulic iron production workshops known as mountainside ironworks (ferrerías de monte) or haizeolak. It has catalogued 170 archaeometallurgical sites of this type in Biscay, and the archeological excavation of some of them has led to the knowledge of the different stages of the production process that took place in these workshops. The team has also experimentally reproduced the work carried out in a mountainside ironworks from the High Middle Ages, and it has currently started the characterization process of this type of activity through the study of the collected materials.     

Mechanism Study on the Wear of CrAlTiN Coated High-Speed Steel Twist Drills Under Dry Cutting Conditions

As demands about environment protection are growing up, dry cutting technology is getting more and more concerns from all over the world. Main works performed here are study on dry cutting performances and wear mechanisms of M2high-speed steel (HSS) twist drills with CrAlTiN multicomponent coatings, which was deposited using magnetron sputter ion plating system, in drilling 45 and 30CrMnSiA steel, and their comparisons to those in drilling the same steel but using monolayer CrN and commercial TiN coated drills. Drilling performances of drills are evaluated mainly through the measurements of width on outer corner flank land and the cutting forces. Results show that performances of CrAlTiN coated drills are better than those of monolayer CrN and TiN coated drills. In drilling 45 and 30CrMnSiA steel, the average tool life of the CrAlTiN coated drills are 17.2 and 11.8 times higher than those of the uncoated drills. Observing wear of the drill with scanning electronic microscope, results show that there is no crack or spallation of the CrAlTiN coatings in wear zones. Main mechanism of the wear here is adhesion.     

Study on Bond Ability of Arc-Spraying Coatings with Different Surface Pretreatment on Cast-Iron

Arc spraying coatings are widely used in various applications, but uncommon in cast iron substrate. Different surface pretreatment technology is tested on substrates of gray cast iron. Surface roughness and residual stress were measured by TR200 and X-ray diffraction analyzer. Influence of different surface pretreatment methods ( dry blasting and fusebond) on roughness and residual stress was analyzed. The arc-sprayed coatings of wire 3Cr13 (φ2mm) on gray cast iron substrate is studied. The microstructure and interface of bonding layer were observed by SEM. The bond strength was taken by tensile test. Results show that bond strength with grit blasting is higher than fuse-bond; it is feasible to make wire 3Cr13 coating with arc spraying on cast iron substrate roughened by grit blasting.     

Structural Characteristics of TiO2 Ceramic Coating by Micro-Plasma Oxidation

TiO2 ceramic coatings with thickness of 20 μm were formed on the surface of pure titanium by micro-plasma oxidation. Their micro-structures were investigated by by using X-ray diffraction and their surface images were detected by using scan electronic microscope. There were three kinds of TiO2 coatings, pure anatase type TiO2 phase, mixed phases consisted of rutile type TiO2 phase and anatase type TiO2 phase, pure rutile type TiO2 phase. The coating surface with the pure anatase type TiO2 phase is rough, while the coating surface with the pure rutile type TiO2phase is smooth. The upper coating surface with the mixed type TiO2 phases is anatase type TiO2 structure and the subsurface of the TiO2 coating is rutile type TiO2structure.     

Evaluation of fatigue life of steel using steel grain size

The insightful discussion of the relationship between the construction of metallographic and fatigue life and proposition of mathematical relationships describing this relationship was conducted in the paper. This paper presents a method for estimating the fatigue life, based on the construction of the microstructural material taking into account the grain size and the participation of ferrite and pearlite phase.