鲮生长激素cDNA的克隆及其重组表达产物的促生长活性

利用RT PCR技术从鲮脑垂体组织中克隆出生长激素cDNA片段 ,克隆的mcGH全长 5 6 7bp ,编码由 188个氨基酸残基组成的GH成熟肽。构建了表达载体pQE30 mcGH ,转化大肠杆菌M15 (pREP4 ) ,在初步优化发酵条件的基础上实现了鲮mcGH在大肠杆菌中的高效表达 ,表达的重组蛋白占菌体总蛋白的 32 13% ,经westernblotting分析证实所表达的重组蛋白是mcGH。表达产物主要以包涵体的形式存在。重组蛋白变性、复性后 ,经Ni chelatingSepharose亲和层析纯化 ,SDS PAGE显示 ,纯化的mcGH纯度约为 94 %。纯化的mcGH腹腔注射莫桑比克罗非鱼 (Oreochromismossambicus) ,经 4周处理后 ,实验组 1和 2的体长增长率分别比对照组快 9 38%和 9 75 % ,体重增长率分别快 2 3 4 2 %和 6 2 4 8%。经t检验统计分析表明 ,表达的重组mcGH具有显著的促罗非鱼生长作用 (P <0 0 5 )。     

重组人IL-6/IL-2融合蛋白的高密度发酵和纯化

克隆的IL-6/IL-2融合蛋白编码基因插入pBV220载体,转化BL21(DE3)菌株中表达。发酵过程中,30℃扩增生长6小时,42℃诱导培养4小时,控制溶解氧在30％～50％。发酵液中最终菌体密度(OD600)达29.17(相当于每升发酵液含55克湿菌体),表达的融合蛋白呈包涵体形式。表达蛋白占菌体总蛋白的30％左右。菌体经过超声破碎后反复洗涤包涵体,融合蛋白纯度达到70％,进一步用离子交换层析和凝胶过滤层析纯化使其纯度达95％以上。IL-6/2融合蛋白中的IL-6和IL-2活性分别为1×107和2×105 U/mg。重组人IL-6,/IL-2融合蛋白在大肠杆菌中达到了中试水平的高表达。     

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。     

苏云金芽孢杆菌营养期杀虫蛋白基因vip3A的研究

根据已知vip3A基因序列设计一对特异性引物VIP3／VIP5,对218株Bt菌株进行PCR鉴定,有51株含有vip3A类基因,其中12株为不同亚种的标准菌株,有4株标准菌株不含有该基因。从Bt C9菌株中分离克隆了vip—C9基因,并插入表达载体pET—21b,转化大肠杆菌BL21,诱导表达出分子量88．6kDa的可溶性蛋白,表达产物对甜菜夜蛾和棉铃虫具有较高的杀虫活性,LC50分别为0．42ng／mg和25．95ng／mg,对小菜蛾活性较低。构建了缺失蛋白Vip—C9—N(N端去除39个氨基酸组成的信号肽序列),杀虫活性测定结果表明其对甜菜夜蛾的活性显著降低,说明N端39个氨基酸对甜菜夜蛾的活性是必需的。     

人肥胖基因在大肠杆菌中的表达

肥胖基因编码的瘦蛋白可以反映机体脂肪含量信息,在发育、繁殖、造血及体重调节等方面具有重要功能。本文利用PCR方法扩增去除信号肽的人肥胖基因cDNA序列,并将位于基因5’端的CCC转变为大肠杆菌常用密码子CCG,扩增片段经测序证实后,克隆原核表达载体pT7-7,重组质粒转化大肠杆菌BL21(DE3),经温度诱导,SDS-PAGE电泳可见分子量约为：16kD的特异蛋白表达带,表达量最高占菌体蛋白总含量的45％以上。表达重组蛋白经纯化,注射昆明白小白鼠,小白鼠体重明显下降,说明纯化的重组蛋白具有生物学活性。     

甘薯β-淀粉酶cDNA克隆及其在大肠杆菌中的表达

用RT-PCR技术从甘薯新大紫(Ipornoea batatas Lam cv．Xindazi)块根总RNA中扩增出预期大小的DNA片段(约1．5kb)。将该片段克隆至pMD—T载体,经酶切分析和序列测定后,正向插入的重组子命名为pMD—cAmy,插入片段长1521bp,编码499个氨基酸残基的多肽,分子量55998kD。克隆的β—淀粉酶cDNA与报道的甘薯另一品种Kokei No．14的mRNA有99％的序列相同,与Calystegia sepiumβ—淀粉酶mRNA有87％相同。重组子pMD—cAmy经IPTG诱导后,胞内可溶性提取物经凝胶电泳和活性染色,出现明显的淀粉酶活性带。SDS—PAGE表明,大肠杆菌表达的β-淀粉酶亚基的分子量约为50kD。重组子pMD—cAmy在淀粉平板上形成明显的水解圈,说明甘薯β-淀粉酶cDNA不仅能在大肠杆菌表达,而且能将有活性的β-淀粉酶分泌到胞外。对重组子pMD—cAmy和甘薯块根可溶性提取物的比较发现,大肠杆菌表达的β-淀粉酶与甘薯块根的β-淀粉酶除迁移率不同外,对淀粉酶活性抑制剂EDTA和β-巯基乙醇的敏感性相同。     

人重组血管生成素在大肠杆菌的表达与鉴定

利用ＰＣＲ技术从人肝细胞文库中扩增出血管生成素的ｃＤＮＡ，将其连入大肠杆菌表达载体后，在大肠杆菌成功地表达出人重组的Ａｎｓ融合蛋白和Ａｎｇ非融合蛋白，表达量占体蛋白的２５％，有明显的促进新生血管生成作用和ＲＮＡ酶及Ｌ９２９细胞粘附活性。     

离子强度对于牛精蛋白在大肠杆菌中的表达的影响

通过基因合成将大肠杆菌偏好的精氨酸密码子CGT取代野生型牛精蛋白基因中的稀有精氨酸密码子AGA和AGG，得到密码子优化的牛精蛋白基因，并将其克隆入原核表达载体pGEX-2T中，组装成表达载体pGEX-2T-PS。将这个表达载体转化入大肠杆菌表达菌株BL21中，经IPTG诱导，可得到一33kD融合蛋白表达带，首次成功地将牛精蛋白在大肠杆菌中进行了表达，但其表达量很低，约为总菌体蛋白的13％。增加表遂菌株培养液的离子强度，可将蛋白表达含量提高到28％，且蛋白表达量与离子强度呈正相关。纯化后的牛精蛋白可在体外条件下与DNA发生结合。     

中国食品包装机械行业产值平稳增长

据报道,今年1～5月,中国食品和包装机械制造行业工业总产值243．23亿元人民币,工业销售产值232．64亿元人民币,同比分别增长9．07％和7．36％。但新产品产值和出口交货值出现下降,分别为8．75亿元人民币、16．5亿元人民币,同比分别下降4．89％和15．82％。     

黄海海燕酸性粘多糖的分离、纯化及鉴定

利用碱提法从黄海海燕中提取了多糖,粗品中糖和蛋白含量分别为28．6％和50．8％;Sevag法脱蛋白后,糖含量57．1％,蛋白含量13．8％;二次酶解后,糖和蛋白含量分别为63．2％和7．0％;粗品除蛋白、脱色后,用25％、42％、66％、88％乙醇分级沉淀分别得到多糖Ⅰ、多糖Ⅱ、多糖Ⅲ和多糖Ⅳ,糖含量分别为20．6％、20．7％、29．2％、19．9％,蛋白含量分别为29．2％、27．8％、59．5％、65．7％;多糖Ⅰ依次用离子交换层析（DEAE-52）和葡聚糖凝胶（G-75）进行分离、纯化,结果表明分子中带有同一种电荷,硫酸基离子鉴定反应为阳性,并至少含有三个组分;用葡聚糖凝胶（G-100）对其分子量进行测定,其平均分子量约为10kD;琼脂糖凝胶电泳结果表明,该糖带负电。研究结果为海燕资源的开发利用提供了科学依据。     

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.     

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.     

Investigation of the Corrosion Behaviors of HVOF-Sprayed Carbide Cernet Coatings in Molten Al-Zn-Si Alloy Bath

In continuous hot-dip galvanization process the corrosion and chemical stability of the sink roll in the galvanizing bath are important problem which effects on the quality and productivity. In order to protect the sink roll the carbide cermet and/or ceramic coatings were deposited on the surface of the sink roll. The WC-, Cr3 C2-cermet coatings were deposited by high velocity oxygen fuel (HVOF) spray, respectively. The coating samples were immersed in molten Zn-alloy containing 50 wt % aluminum at 833 K for 24 hr and 144 hr, respectively. The inter-diffusion and inter-reaction of Zn, Al and elements in coating and corrosion behaviors of these coatings were investigated by XRD, SEM and EPMA etc. The corrosion mechanisms of the carbide cermet coatings and ceramic coatings in molten High Al-Zn-alloy were approached.     

Fundamental study of adhesion of ice to cooling solid surface (Discussion on influence of copper oxide layer on ice adhesion force)

In many situations, ice often adheres to a cooling solid surface, frequently causing serious accidents. It is critical to clarify the mechanism of ice adhesion to the cooling surface in order to prevent ice adhesion. In a past study, the shearing stresses of two kinds of test plates with a copper surface having the higher thermal conductivity were measured. The shearing stress corresponds to ice adhesion force. Both shearing stresses were significantly different; however, the cause remains unclear.Therefore, the present study focuses on an oxide layer as the main factor causing the difference of both shearing stresses; the influence of the oxide layer formed on shearing stress was discussed. And in the removal and reformation processes of the oxide layer, the time variation of the shearing stress was clarified. Moreover, the relationship between the state of the copper surface and the shearing stress was also clarified by surface analysis.     

重塑标准推行格局 推动LED照明的普及（英文）

正When you are enjoying the green,energy-saving and healthy lighting of IKEA LED luminaire,you may not realize that most of IKEA’s LED bulbs are produced by the LED luminaire smart manufacturing system of Hangzhou Zhongwei Photoelectricity CO.,LTD(ZVISION).No longer enclosed in the laboratory,the product standards are brought into the factory workshop and even to the entire production process.Thus,the standards are promoted and applied in a favorable way by enterprises,benefitting consumers with high quality products.     

Chinese Experts Contribute to ISO＇s New Toy Standards

正The international standard,ISO 8124-6,Safety of toys–Part 6:Certain phthalate esters in toys and children’s products,was officially issued by ISO worldwide on August 15.It was the first time for Chinese experts to lead the ISO standard development in toys industry.The standard was drafted by a group of Chinese experts from SAC/TC for toys and relevant Chinese departments and finally completed by a dedicated international group of experts.ISO 8124-6 makes a substantive breakthrough in the aspects of the scope of toys,limitation of plasticizer types,control of test cost,accuracy of test,operation convenience,etc.,offering the operational guidance for toy manufactures and testing laboratories.     

中国质量(北京)大会(英文)

正Themed on"quality,innovation,development",the first China Quality Conference(Beijing)was held in the Great Hall of the People on September 15.Chinese Premier Li Keqiang delivered an important speech at the conference.State Councilor Yang Jing and Wang Yong attended the conference with over 600 domestic and international representatives from foreign,regional and international quality organizations     

National Working Meeting on Land and Resources Standardization

正The National Working Meeting on Land and Resources Standardization was held in Beijing on July 15,2014.Wang Min,the Deputy-Minister of the Ministry of Land and Resources,and Yin Minghan,the Chief-Engineer of SAC addressed the meeting.Representatives from relevant competent departments and technical committees were present.The meeting aims to summarize work,discuss existing problems and make clear working thoughts,promoting standardization work of land and resources to a new level.Wang Min pointed out that the standardization work of land and resources should be centered on the basic principles of"protecting land and resources dutifully,intensively and economically utilizing land and resources,and making all-out efforts to safeguard the public’s rights and interests",highlighted the