再纯化

Abortus, amplify BP26 gene by PCR and clone to pMD18-T simple vector. Identify the constructed recombinant plasmid pMDBP26 by sequencing, then subclone to vector pET-28a. Transform the constructed recombinant plasmid pETBP26 to E. coli BL21(DE3) for expression under induction of IPTG. Purify the expressed product by histidine-binding resin column chromatography and identify by Western blot.

提取布鲁氏菌基因组DNA，用PCR扩增出BP26基因，并克隆到pMD18-T载体上，测序正确后，再亚克隆至pET-28a载体上，转化到大肠杆菌BL21(DE3)中，用IPTG诱导表达，经组氨酸结合树脂柱纯化，并对纯化后的表达产物进行Western blot鉴定。

In this study, we isolate, purify and characterize endo-β-glucanase by Ascomycete Xylaria regalis. The cellulose-containing medium is used to induce the production of the cellulose-degrading enzymes. When these enzymes are produced at maximum level, we harvest and filtrate the culture medium. The filtrate is purified by anion exchanger column and hydrophobic interaction column (phenyl-650M). The purification fold of endo-β- glucanase increases 4.3 times, the specific activity approaches 3.7 unit/mg, and the recovery is 27.6%, respectively.

为了分离且纯化此株菌产生在培养基中的endo-β-glucanase，先将此菌在cellulose液态培养基中大量培养，在此酵素达到最高峰时，利用滤纸过滤将菌丝及菌液分开，收集粗酵素液，再经过阴离子交换树脂及疏水性层析管柱(Phenyl-650M)等步骤予以纯化，所得到最终酵素纯度为 4.3倍，比活性为3.7 unit/mg，回收率为27.6%。

However, CDH has little synergism with cellobiohydrolase or endoglucanase in cellulose degradation. But, if the substrate was pretreated with CDH in the presence of cellobiose, the rate of the hydrolysis of the substrate by cellobiohydrolase or endoglucanase increased.

CDH与纯化的外切纤维素酶或内切纤维素酶或内切纤维素酶之间没有协同作用关系，但在纤维二糖存在条件下底物事先用CDH处理后再用上述纯化的酶水解，作用明显增强。

This paper develops the method of extraction and purification of thrombin from the porcine plasma by using Ion Exchange.

对离子交换法提取纯化凝血酶的方法进行了改进：首先在-20℃对猪血浆进行冷冻处理，然后再采用离子交换法提取纯化凝血酶。

First of all, this study will be multi-walled carbon nanotubes by chemical oxidation process purified so it can be modified to deal with than the purification of purity before the modified 5 wt%, more than 1.89 times the surface area to increase the surface potential decrease of about 10 ~ 20 mV and the surface functional base (-COOH and-OH) to increase 1.45 times, and then the control technology of electroless preparation parameters include: analysis of plating time, plating analysis of temperature, metal ions in solution than the (Fe2+/ Ni2+), bath pH and the added value of dispersion Agent and other research towards Fe-Ni particles to increase the iron content and Fe-Ni particles spread in the MWCNT upper fixed targets,the results found that when the parameters for the preparation of 50 ℃, pH10, do not add dispersant, metal ions than the solution (Fe2 +/ Ni2 +) 7, the highest iron content can be Fe = 40.55 at%(flat iron content 111.17 mg / g) of nanocomposites Fe-Ni/CNT, then for a series of the nature of its analysis, the final evaluation Fe-Ni/CNT nanocomposites were processed on the effectiveness of the application of mixed pollutants, the results showed that in 120 minutes at the same time when Adsorption of heavy metal ions lead nitrate 10 ppm, selenium heavy metal ion degradation of 1 ppm and orange azo dye AO7 50 ppm standard of effluents, and after a total Fe-Ni nano-particles to SEM / EDS analysis of more than 50% still remaining.

本研究首先将多壁奈米碳管以化学氧化法做纯化改质处理能使其纯度较纯化改质前提高5 wt％、比表面积提高1.89倍、表面电位下降约10~20 mV以及表面官能基提高1.45倍，再控制无电镀技术之制备参数包括：析镀时间、析镀温度，镀液中金属离子比(Fe2+/Ni2+)、镀液pH值及是否添加分散剂等，研究朝提高Fe-Ni粒子的含铁量与Fe-Ni粒子散布固定在MWCNT上等目标进行，研究结果发现当制备参数为50 ℃、pH10、不添加分散剂、镀液金属离子比(Fe2+/Ni2+)7时，可得到最高铁含量Fe = 40.55 at％(单位铁含量111.17 mg/g)之奈米复合材料Fe-Ni/CNT，接著对其作一系列性质分析，最后评估奈米复合材料Fe-Ni/CNT对共处理混合污染物之应用效益，结果显示其在120分钟时能够同时吸附重金属硝酸铅离子10 ppm、重金属硒酸根离子1 ppm及降解偶氮橘色染料AO7 50 ppm达放流水标准，且共处理后奈米Fe-Ni粒子以SEM/EDS分析仍剩余50 %以上。

It was further purified by ultracentrifuge and repeated precipitation and purification with ethanol to obtain the required polysaccharide components, in which MEP\|SP of molecular of 10 000～100 000 was purified and separated by...

对其中分子量为 10 0 0 0～ 10 0 0 0 0的多糖，采用DEAE 纤维素离子交换柱，以不同浓度NaCl为洗脱液，进行纯化分离，再经SepharoseCL 6B纯化得到 2种水溶性多糖MEP SP2和MEP SP3。

Coli genome was amplified by using PCR method. The PCR product was cloned into PUC19-T vector and sequenced. In addition, the Mn-SOD protein expressed by pET-28a vector and purified was injected into rabbits to prepare polyclonal antibody. Western-blot was used to detect the effect of polyclonal antibody.

coli基因组中扩增Mn-SOD基因编码区，将它克隆到原核表达载体上进行大量表达和纯化，再用纯化的蛋白对新西兰大白兔进行背部多点注射，40d后取其血清，用Western-blot印迹实验测定抗体效果。

Chemiluminescent immunoassay is one of the advanced immunoassay ofnon-radioisotopic immunoassay because of its high sensitivity, wide dynamic range,high accuracy, stable labeled protein, full automation and extensive application field.CLIA using acridinium ester derivatives as chemiluminescent label has advantages oflow background, high signal-to-noise ratio, no need of catalyst and simpleluminescence system. Different aspects associating the DMAE·NHS-based CLIA were studied in thisthesis, including synthesis of acridinium ester, chemiluminescentcharacteristics of DMAE·NHS, labelling antibody or streptavidin with DMAE·NHS,two-site sandwich chemiluminescent immunoassay for TSH, two-sitesandwich chemiluminescent immunoassay involved biotin-streptavidinsystem for TSH, and competitive chemiluminescent immunoassay usingbiotin-streptavidin system for TT〓. The desired acridinium ester, DMAE·NHS, was synthesized according to thereference method with some modifications. The products were identified by IR, NMR,MS and elemental analysis. In our method, KOH was used in place of NaOH tosynthesize benzyl ester of 3,5-dimethyl-4-hydroxybenzoic acid,2',6'-dimethyl-4'-(n-succinimidyloxycarbonyl) phenyl-acridinium-9-carboxylate waspurified on a silica gel column with chloroform/ethylacetate(4:1, v/v) as eluent andfurther purified by triturating with hexane/acetone(2:1, v/v).

DMAE·NHS的合成是本论文的关键和难点，我们对文献方法进行改进：文献方法用氢氧化钠与3，5-二甲基-4-羟基苯甲酸反应制得3，5-二甲基-4-羟基苯甲酸钠，再用3，5-二甲基-4-羟基苯甲酸钠与苄氯作用制备3，5-二甲基-4-羟基苯甲酸苄酯，我们用氢氧化钾代替氢氧化钠，使合成取得成功；在合成2'，6'-二甲基-4'-苯基-吖啶-9-甲酸酯时，文献方法对粗产品进行两次硅胶柱层析纯化，第一次柱层析用氯仿／乙酸乙酯(4∶1，V∶V)作溶剂和淋洗剂，第二次柱层析用己烷／丙酮(2∶1，V∶V)作溶剂和淋洗剂，按照文献方法得到的不是所需要的化合物，我们只进行第一次柱层析纯化，然后用己烷／丙酮(2∶1，V∶V)进行研磨，过滤，洗涤，除去溶于己烷／丙酮(2∶1，V∶V)的部分，得到所需要的产品。

The method to purify compound 4 was resolved. A novel method to prepare the intermediate from phenylacetyl chloride was selected in this paper. The facile synthetic scheme put forward in this work is simple to operate and has the characteristic of high yield and good feasibility.

解决了终产品4的分离纯化难题，采用生成衍生物再进行水解的方法达到了纯化目的；提出了合成中间体3-羟基-4-苯基-吡喃-2，5-二酮(7)的较为新颖的方法，操作简单，各步中间体无需作进一步纯化便可进行下一步反应，从而得到了结构较新颖的终产品4。

Dorsalis by salting out, ion exchange chromatography, and gel filtration chromatography. By salting out the egg homogenate, SDS-PAGE analysis revealed that the Vns were mainly precipitated in a 50~70% ammonium sulfate solution. The salted-out proteins were further purified by ion exchange chromatography, and the result showed that Vns were mainly eluted with a 0.3~0.5 M sodium chloride gradient. Finally, the elutes of ion exchange chromatography were refined by running them through gel filtration chromatography to obtain purified Vns. SDS-PAGE separated the 48- and 51-kDa purified Vns, and used them as antigens to raise polyclonal antibodies.

利用硫酸铵盐析法初步分离果实蝇卵中的蛋白质，以SDS-PAGE蛋白质电泳分析得知，卵黄蛋白主要在硫酸铵溶液浓度为50~70%时被沉淀出；盐析沉淀所得之卵黄蛋白，再以离子交换层析法进一步纯化显示，卵黄蛋白主要会在0.3~0.5 M氯化钠的浓度范围间被析出来；再将析出液浓缩后，经凝胶过滤层析法做最后的纯化，即获得纯化之卵黄蛋白。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失