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A multiplex PCR assay for detection of Escherichia coli O157:H7 was developed by using 3 sets of primers that specifically amplify segments of the rfbE、fliC and eaeA genes. The target genes fragment of the PCR assay were 291 bp, 625 bp and 368 bp, respectively. Analysis of 30 strains demonstrated that this PCR system was specific.

以编码大肠杆菌O157抗原的rfbE基因、编码H7抗原的fliC基因以及编码毒力因子的eaeA基因为靶基因,选择3对引物,建立并优化了检测大肠杆菌O157:H7的多重PCR体系,扩增产物分别为291 bp、625 bp、368 bp,采用30株细菌验证了该多重PCR具有特异性。

The flic gene of App was obtained by PCR based on highly homology with Escherichia coli, Salmonella, Shigellosis. Then it was legated to pMD-18T vector. The recombinant plasmid was identified by enzyme and sequence analysis. The 528 bp fragment of flic gene was successfully cloned into pGEX-KG vector, then the recombinant plasmid was transformed into E.

以猪传染性胸膜肺炎放线杆菌血清Ⅰ型菌株基因组为模板,根据其鞭毛区与其它细菌高度同源性设计引物,利用PCR方法扩增鞭毛蛋白基因flic片段,将其亚克隆到pMD-18T载体中并进行PCR和酶切鉴定,再将其亚克隆与载体pGEx-kG分别双酶切和连接,构建重组表达载体pGEX-flic,并将其转化大肠杆菌工程菌BL21,进行原核表达。

Four vectors of pET vector system were chosen to express the enzyme in E. coli BL21(DE3). The main results were as follows: 1. The total RNA was obtained from fungus Syncephalastrum racemosum.

以总状共头霉菌株总RNA为模板,通过RT-PCR方法,获得了去除自身信号肽的天冬氨酸类蛋白酶syncephapepsin基因。2。

The radical scavenging activity of Laminaria japonica pigments was studied by the DPPH2,2-Diphenyl~(-1-picrylhydrazylassay. The total pigments showed a strong DPPH radical scavenging activity of 65.819% with the concentration of 0.52mg.ml~(-1). The DPPH radical scavenging activity of fucoxanthin was 19.85% with the concentration of 0.27mg.ml~(-1). The antimicrobial activity of pigment in Laminaria japonica was studied by the method of round paper. The results showed that the pigment extracts have activity against Escherichia coli, Pseudomonas aeruginosa, Saricina flava and Saccharomyes cerevisia. The antibacterial activity of total pigments for Saccharomyes cerevisia was the strongest with an inhibition diameter of 18.9 mm; and the antibacterial activity of fucoxanthin for Saricina flava was also much significant with an inhibition diameter of 10.2 mm.

测定了它们对自由基的清除率和对四种细菌的抑菌效果,研究结果表明,总色素和褐藻黄素的浓度与自由基清除率成正比,当总色素浓度为0.52mg.ml~(-1)时,清除率达到65.819%,高于陆生植物黑米色素和紫甘薯色素;当褐藻黄素浓度为0.27mg.ml~(-1)时,自由基清除率达到19.85%,低于紫苏色素、紫甘蓝色素、黑米色素、紫甘薯色素;对大肠杆菌、绿脓杆菌、八叠球菌、金黄葡萄球菌的抑菌实验结果显示,海带总色素对金黄葡萄球菌的抑菌活性最强,抑菌圈直径为18.9mm;褐藻黄素对八叠球菌的抑菌活性相对较强,抑菌圈直径为10.2mm。

After optimizing the expression conditions, functional xylanase XT6 was over expressed in E. coli with up to 65% of total protein.

通过优化表达条件,功能正常的XT6基因在大肠杆菌中成功过量表达,蛋白表达量占细胞中总蛋白的65%。

Lividans. Using pIJ903 bearing tsr gene as a vector, the two furthermost fragments of the cluster, 4. 0kb and 1. 5kb in size respectively, were inserted into it. To facilitate the detection of gene replacement, apr gene was placed in the middle of the two inserts. OriT from E coli plasmid RK2 was also incoporated into the vector, therefore, pIJ903 derivatives can be mobilized from E.

在具有硫链丝菌素抗性基因的pIJ903的单—BamHI位点,同向插入了FR-008生物合成基因簇两个最远端的4.0kb和1.5kb片段,并在二者之间插入了可在链霉菌FR-008和变铅青链霉菌中表达的阿泊拉霉素抗性基因,同时也插入了有助于将质粒引入链霉菌的oriT片段,从而构建出了置换整个基因簇的基因置换质粒pHZ691。

Coli mainly in a form of inclusion body. The expressed product contained about 25% of total somatic protein and showed good reactogenicity as proved by Western blot. The IgG level against GAMP induced by GAMP-CRM197 conjugate was significantly higher than those by GAMP and by the mixture of CAMP and rCRM197. Repeated immunization with the conjugate induced immunopotentiation, indicating the effect of CRM197 as a carrier protein.

结果重组CRM197在大肠杆菌中主要以包涵体形式表达,表达量占菌体总蛋白的25%左右;Western blot证明重组rCRM197具有良好的反应原性;各针接种后,结合物诱生的多糖特异性IgG水平均显著高于GAMP组和GAMP+rCRM197混合物组,具有较强的免疫原性;多次接种产生了免疫增强效应,重组CRM197具有载体蛋白的作用。

In this paper, a new product of Ce/ TiO2 compound antibacterial agent has been made, comparing with naked nano-TiO2, it has better antimicrobial activity against E.coli ,S.aureus and scorch gemmule bacillus in the solar light, increasing the quantity of Ce4+, exhibiting stronger antibacterial property.

制备出稀土铈掺杂锐钛型纳米TiO2复合抗菌功能材料,该抗菌材料与不掺杂的纳米TiO2相比,在可见光下对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌的混合菌具有较好的抑菌效果,随着Ce4+的摩尔浓度增加,制备的试样的抗菌能力越强。

The above experiments showed that the UHP has an impact on E.coli DH5αgerm plasm.

通过以上实验,说明超高压对大肠杆菌遗传物质产生了影响。

The expression vector pET32a-ITF was constructed and expressed in E.coli successfully, and the recombinant protein rITF and its antibody was also purified in this study. ITF expressed in small intestine goblet cell.

成功构建了表达载体pET32a-ITF,在大肠杆菌中表达并纯化获得纯度较高的rITF,并获得了生物活性较高的ITF抗体,ITF主要在肠道杯状细胞分泌表达。

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