HIV-1中国流行株gp120基因在痘苗病毒中的表达及其与核酸疫苗的实验免疫研究

将HIV－1中国流行株gp120基因在痘苗病毒中进行表达,以期获得重组痘苗病毒,与核酸疫苗混合免疫,评价免疫效果,为艾滋病疫苗开发研制打下基础。将HIV－1中国流行株gp120基因片段插入到pJ38载体启动子下游,经同源重组和血凝素阴性空斑筛选重组痘苗病毒,SDS－PAGE、Western blot检测目的蛋白。以重组病毒和核酸疫苗免疫BALB／c小鼠,进行淋巴细胞转化实验、CTL、CD4＋CD8＋T细胞数目以及血清抗体等细胞免疫和体液免疫指标检测。结果获得的重组痘苗病毒vJ38pg120能够表达Gp120蛋白并诱导机体产生细胞免疫和体液免疫,具有良好的免疫原性,其免疫效果以2rVV-DNA混合方式为最好。重组痘苗病毒vJ38gp120。     

人免疫缺陷病毒I型p24gag基因的重组与表达

将编码人Ｉ型免疫缺陷病毒（ＨＩＶ－１）衣壳蛋白的ｐ２４ｇａｇ基因片段，克隆到原核表达载体ｐＥＴ－１７ｂ的Ｔ７噬菌体启动子下游，构建成了重组表达质粒ｐＥＴ２４，并使ｐ２４ｇａｇ基因片段在大肠杆菌ＢＬ２１（ＤＥ３）中高效表达，产物为３０ｋＤａ的ｓ１９－ｐ２４融合蛋白，表达量占菌体总蛋白的３８．４％。重组ｐ２４蛋白均与抗ｐ２４单克隆抗体及ＨＩＶ－１阳性血清发生特异性反应，具有较好的抗原性。     

鸡痘病毒282E4株基因组末端非必需区的序列测定与分析

将鸡痘病毒（ｆｏｗｌｐｏｘ ｖｉｒｕｓ，ＦＰＶ）２８２Ｅ４株７．３ｋｂ的ＢａｍＨＩ片段经亚克隆后，获得ｐＵＦａ，ｐＫＳＦｂ，ｐＵＦｃ和ｐＵＦｄ四个重组策粒。采用ＡＢＩ ３７７ＤＮＡ测序仪荧光标记法对上述质粒进行了序列测定，并应用ＤＮＡ ｓｉｓ Ｖ７．０同ＧｅｎＢａｎｋ中已知的ＦＰＶ序列和痘苗病毒的Ｃｏｐｅｎｈａｇｅｎ株的全序列进行同源性比较。     

人骨形态发生蛋白－1（hBMP－1）成熟肽的基因克隆及其在大肠杆菌中的表达

采用ＰＣＲ方法从ＢＭＰ－１的ｃＤＮＡ中克隆了ＢＭＰ－１成熟肽的编码基因，删除了ＢＭＰ－１前体分子Ｎ端的信号肽序列和Ｃ端的其他序列。用ＨｉｎｄⅢ消化１．３Ｋｂ的ＰＣＲ产物，将０．８４和０．４６Ｋｂ两片段分别克隆到ｐＵＣ载体中进ＤＮＡ序列分析。分别酶切回收ＥｃｏＲ－ＨｉｎｄⅢ和ＨｉｎｄⅢ－ＢａｍＨⅠ两目的基因片段，与大肠杆菌表达载体ｐＢＶ－２２０进行退火连接，使插入基因受控于Ｐ＿ＲＰ＿Ｌ启动子。将含有ＢＭＰ－１成熟肽编码基因重组表达质粒ｐＢＶＢＭＰ－１转化大肠杆菌宿主细胞进行温度诱导表达。结果表明，经４２℃热诱导后，大肠杆菌能以包涵体形式表达ＢＭＰ－１成熟肽，其分子量约为５２ｋＤａ。     

共表达H5、H7亚型AIV HA基因与鸡IL-18基因的重组鸡痘病毒构建

以鸡痘病毒(FPV)疫苗株为载体,将H5和H7亚型禽流感病毒血凝素(AIV HA)基因串联后(拥有同一个阅读框)和鸡白细胞介素-18(IL-18)基因分别插入到鸡痘病毒表达载体pUTA-16-LacZ复合启动子(ATI-P7.5×20)和单一启动子1(P7.5)下游,构建了携带AIV HA基因和鸡白细胞介素-18基因的重组鸡痘病毒转移载体质粒pUTAL-H5-H7-IL18;用相同的方法构建重组鸡痘病毒转移载体质粒pUTAL-H5-IL18;将H5亚型AIV HA基因插入到鸡痘病毒表达载体pUTA2复合启动子(ATI-P7.5×20)下游,构建了携带H5亚型AIV HA基因的重组鸡痘病毒转移载体质粒pUTA2-H5.应用脂质体转染法,将重组鸡痘病毒转移载体质粒与282E4株鸡痘病毒共转染鸡胚成纤维细胞(CEF),经BrdU进行三次加压蚀斑筛选后,以不同代次的细胞mRNA为模板,利用H5亚型AIV HA基因、H7亚型AIV HA基因和鸡IL-18基因特异引物进行RT-PCR和蛋白印迹检测,筛选出H5HA-H7HA融合蛋白基因和鸡IL-18基因共表达的重组鸡痘病毒rFPV-H5HA-H7HA-IL18,H5亚型AIV HA基因和鸡IL-18基因共表达的重组鸡痘病毒rFPV-H5HA-IL18以及单独表达H5亚型AIV HA基因的重组鸡痘病毒rFPV-H5HA.这些重组鸡痘病毒的构建为AIV活载体疫苗的研制奠定了基础.     

荧光素酶报告基因在转基因蚕中的应用

报道了在针对ＢｍＮＰＶ的核酶转基因蚕筛选过程中利用荧光素酶报告基因的检测及鉴定结果。试验采用的质粒为含家蚕核多角体病毒的即刻早期蛋白基因ＩＥ启动子－荧光素酶报告基因－抗ＨＰＶ ＩＥ核酶基因的联合重组质粒ｐＩＥ－Ｌｕｃ－Ｒｚ。该质粒经限制性内切酶Ｓｃａ Ｉ线性化后用基因枪法导入家蚕早期受精卵,待Ｇ１代测定荧光素酶活性表达较高的蚕的羽化交配产卵,分别于Ｇ２,Ｇ３和Ｇ４代再用ＢｍＮＰＶ接种筛选出对ＮＰＶ     

抗CD3基因工程嵌合抗体片段Fab‘的构建,表达和活性测定

ＰＣＲ扩增抗ＣＤ３单抗轻链可变区和重链可变片段基因，将其重组到Ｆａｂ’表达载体中，构建成抗ＣＤ３嵌合抗体Ｆａｂ’表达载体，转化大肠杆菌１６９Ｃ９进行可溶性表达。产物经蛋白Ｇ亲和层析柱纯化。免疫荧光竞争结合实验和＾３Ｈ反故实验证实能与小鼠抗ＣＤ３ＩｇＧＨＩＴ３ａ竞争性结合表达ＣＤ３的Ｔ淋巴细胞，并促进细胞增殖。     

HIV-1核心蛋白Gag与IL-6共表达重组鸡痘病毒的构建及其免疫原性研究

分别将IL-6和HIV-1gag基因插入到鸡痘病毒表达载体质粒pUTAL串联启动子和复合启动子(ATI-P7．5)下游，构建了重组鸡痘病毒表达质粒pUTA-GAG-IL6。经同源重组和Western blot检测，获得重组鸡痘病毒，并将该重组病毒免疫BALB／c小鼠，检测免疫小鼠血清抗体水平和特异性CTL杀伤活性。结果获得的重组鸡痘病毒可同时表达HIV-1核心蛋白和IL-6，并在感染细胞内形成病毒样粒子。重组病毒具有良好的免疫原性，可诱导小鼠产生特异性体液免疫和细胞免疫。     

口蹄疫三价重组鸡痘病毒疫苗分子设计及其构建

针对我国口蹄疫(FMD)流行情况,利用计算机软件模拟分析,构建了口蹄疫病毒(FMDV)复合多表位基因工程疫苗表达盒OAAT.该表达盒以O型、A型FMDV结构蛋白VP1全基因和AsiaI型FMDV 2个基因拓扑型的结构蛋白VP1基因上的5个抗原表位基因(其中2个抗原表位来源于FMDVIND63/7株,该毒株与我国新疆分离株属于同一基因拓扑型,其余3个抗原表位来源于FMDVYNBS/58株)作为主要免疫原基因,以非结构蛋白上的2个Th2细胞表位基因及结构蛋白上的1个Th2细胞表位基因作为辅助基因.将构建好的表达盒OAAT和猪IL-18基因分别插入到鸡痘病毒(FPV)表达载体pUTA-16-LacZ复合启动子(ATI-P7.5×20)和单一启动子(P7.5)下游,构建了携带OAAT表达金和猪IL-18基因的重组鸡痘病毒转移载体质粒pUTAL-OAAT-IL18.应用脂质体转染法,将重组鸡痘病毒转移载体质粒与282E4株FPV共转染鸡胚成纤维细胞(CEF),经BrdU进行3次加压蚀斑筛选后,以不同代次的细胞mRNA为模板,利用FMDV VP1基因和猪IL-18基因特异引物进行RT-PCR和IFA检测,筛选出OAAT和猪IL-18基因共表达的重组鸡痘病毒rF-PV-OAAT-IL18,该重组鸡痘病毒的构建为新型FMDV活载体疫苗的研究奠定了基础.     

《高技术通讯》2001年总目录

栏目·篇名作者　期 (页 )……………………………………栏目·篇名作者　期 (页 )……………………………………生物技术小白鼠心肌核ＤＮＡ片段中的基因表达的分子开关的研究袁明秀等 1(7)……对虾白斑杆状病毒 (ＷＳＢＶ)定量ＰＣＲ检测技术研究徐丽美等 1(14 )………农杆菌介导的油菜脂肪酸调控基因工程研究石东乔等 2 (1)………………ＨＩＶ 1中国流行株 ｇｐ12 0基因在痘苗病毒中的表达及其与核酸疫苗的实验免疫研究王莉馨等 2 (8)………………………………基因枪法转化水稻 (ＯｒｙｚａｓａｔｉｖａＬ .)获得可育的转抗虫基因水稻再…     

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.     

Effect of Permeable Crystalline Material on Steel Reinforcement Corrosion of Concrete

Permeable crystalline materialcan permeate into pores and cracks of concrete and catalyze the reaction between Ca(OH) 2and unhydrated cement to generate a great quantity needle non-soluble crystals, which can stop up the pores and cracks of concrete, and increase the impermeability of concrete. This paper reported the results of a study conducted to evaluate steel reinforcement corrosion of concrete specimens uncoated and coated with permeable crystalline material as well as mixed with the permeable crystalline material. The properties evaluated for corrosion test were water impermeability, water absorption, compressive strength and potential. The results of water impermeability, water absorption, compressive strength clearly showed that the permeable crystalline material could prohibit water, any soluble salts and moisture from penetrating the concrete to cause corrosion, leaking, and other problems, and it did increase the compressive strength, which was favorable for protection of corrosion of reinforcing steel. Moreover, it was concluded from the potential-time curve that the steel reinforcement of uncoated specimen was in the state of activation whereas that of other specimens coated and mixed with the permeable crystalline material was in the state of inactivation. Above all, it was indicated that the permeable crystalline materialis very effective to protect the steel reinforcement of concrete from corrosion.     

The Research and Application About "Dry Peeling Off" Technology for Airplane Surface Paint Layer

In this paper, the principle of spurting take off lacquer were introduced; and with orthogonal method the unsaturate polyester was prepared ;it was easy to peel airplane paint layer off with this material and equipment.     

Quality Study of Honeycomb Core Shaping Without Wax Stabilization

Sandwich panels utilizing honeycomb cores result in lightweight, high-strength units that are very rigid. Indeed, they give honeycomb panels the best strength-to-weight ratio of any construction type available. Conventionally, the wax is poured into the cell of the core to strengthen the structure and prevent the core from being crushed during milling. Much time and labor is required to wax, mill, and clean the cores. In this study, the process of pouring wax into the core was skipped. In other words, the cutter directly milled the core. Experiments showed that the compressive strength of the core produced without wax stabilization was only 3% lower than that of cores produced using the traditional method. In addition, the drum peel strengths of cores produced without wax stabilization were 38.4% higher than cores produced using the traditional method, in which residual wax seriously polluted the adhesive. It was also found that eliminating the wax process can reduce costs.     

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.     

In-process thermal treatment of polylactic acid in fused deposition modelling

Polylactic acid (PLA) is one of the most widely used open source fused filament fabrication materials due to its ease of extrusion, biodegradability, and mechanical strength. The mechanical strength of PLA largely depends on the proper growth of its semi-crystalline structure, which can be severely impaired by a low rate of crystallization, particularly in open source printers. This can be further aggravated by the non-uniform thermal distribution of heat that causes improper curing among the extruded beads of the printing material. As a result, PLA printed on open source printers does not achieve the best mechanical properties. This research, for the first time, proposes an additive-free solution implemented through a detailed set of experimentation to improve the curing rate through in-process temperature variations to cure the joints among the beads. Scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy is used to confirm the improvements in the bead joints. This work is conducted in two phases of experiments. In the first phase, a full factorial ANOVA is used to investigate various process parameters and the important variables are used in the second phase to print test specimens in four different sets.