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After transfection of E.coli BL21 and induction by IPTG,the recombinant immunotoxin fusion protein was highly expressed in E.coli BL-21 in the form of inclusion bodies(up to 32.29% of total cellular proteins).

IPTG诱导后目的蛋白获得高效表达，SDS-PAGE分析目的蛋白主要以不溶性包涵体的形式存在于菌体中，表达量占菌体总蛋白的32.29％。

Coli Min27 strain with mitomycin C. Twenty-one E. coli strains with various serotypes were infected with FMin27, and plaque formation and lysogenic conversion of them were investigated.

用丝裂霉素对该突变菌株进行诱导，结合抗性标记和PCR方法筛选出一株突变的志贺毒素2型噬菌体，命名为FMin27。

Results We obtained a lysogenic bacteriophage MZTP01with clear plaque and 1 mm diameter. Fragment D with 2362bp (Genebank No. AY639599) was obtained after the phage DNA hydrolyzed by HindⅢ/EcoRⅠ. Among fragment D, the pep gene with molecular weight of 47 kDa and length of 1101bp was cloned and expressed. Recombinant M15 (pQE30pep) was built and overexpressed in Escherichia coli with a 47kDa clear band. At the same place a clear band was observed by Western blot. Judging from the time course expression, we could conclude that PEP protein produced at 1 hour after induction and then increased gradually. PEP protein was mainly in the form of inclusion body in the recombinant and slowed the growth speed of host. Homologous comparison of PEP protein from phage MZTP01 with other PEPs from BLAST were that phage MZTP01 PEP protein had 100% homologe with that of Escherichia coli K12, and most of others took the similarity in the range between 37%~84%.

诱导获得的溶原性噬菌体MZTP01斑点清晰，直径约1mm，成斑时间12h；从噬菌体基因组DNA双酶切(HindⅢ/EcoRⅠ)片段中回收长度为2362bp的D片段(Genbank登录号： AY639599)，又从D片段中克隆了长度为1101bp、编码367aa、分子量为47kDa的pep基因，表达载体M15(pQE30pep)在大肠杆菌(Escherichia coli， E.coli)中表达获得了47kDa的清晰表达带，在1h 时开始产生蛋白并有逐步上升的趋势； Western blot 也在47kDa处得到一条清晰的条带；可溶性分析表明PEP蛋白在重组菌株中是以不可溶的包含体形式存在的，该蛋白的产生明显地抑制了宿主的生长速度；噬菌体PEP氨基酸序列之间的同源性比较表明，噬菌体MZTP01 PEP蛋白与来自E.coli K12噬菌体的PEP蛋白的同源性程度最大。

Results totally 287 strains were isolated from the 256 positive samples , and the gram-negative bacilli were 225(78.4%,), the gram-positive coccus were 41(14.3%), snd the monilia were 21(7.3%).the distributions of clinical bacteria were respiratory tract(63.4%),urinaryract(7.0%),secretion(includingwound .3%),blood(5.9%),stool(5.2%), pucture fluid(4.9%), and other sites(7.3%). of all isolating bacterium,from the first to the fifth were ps.aeruginosa(19.5%),k.pneumoniae(16.7%), e.coli(14.3%), a.baumannii(11.8%) and psemal (10.1%).resistant rates of methecillin-resistant s.aureus,methecillin-resistant coagulase-negative staphylococci and vancomycin-resistantwere 88.2%、70.0% and 11.1% respectively;the incidence of e.coli and k.penumoniae produce extended speutrum beta-lactamase were 68.6%和65.2%, 44.6% of ps.aeruginosa isolates were resistant to imipenem; the highest examining rate of 21 kinds of monilia was candida albicans (66.7％),resistant rate of candida albicans to fluconazole and amphotricin b was 51.3％ and 1.3%.

结果 在254份检出细菌阳性标本中共培养出287株细菌，其中革兰阴性杆菌225株（78.4％），革兰阳性球菌41株（14.3%），念珠菌21株（7.3%），检出菌来自呼吸道标本占63.4%，其他标本各占5%左右；细菌检出占构成比前三位的依次为铜绿假单胞菌19.5%、肺炎克雷伯菌16.7%、大肠埃希菌14.3%；耐甲氧西林金黄色葡萄球菌、耐甲氧西林凝固酶阴性葡萄球菌和耐万古霉素的肠球菌的发生率分别为88.2%、70.0%和11.1%；大肠埃希菌和肺炎克雷伯菌的超广谱β-内酰胺酶的检出率分别为68.6%和65.2%，铜绿假单胞菌对亚胺培南的耐药率为40.2%；白色念珠菌对氟康唑的耐药率为81.3％，对两性霉素的耐药率为3.2%。

E.coli RNA polymerase can transcribe all the genes of E.coli, but the three eukaryotic RNA polymerases transcribe only specific, nonoverlapping subsets of eukaryotic genes.

eukaryotic genes 大肠杆菌的RNA聚合酶能够转录所有的基因，但真核生物的三种RNA聚合酶各自只能转录特定的、不相重叠的一组基因。

The minimal bactericidal concentration of PTKE on Escherichia coli, Staphylococcus aureus, Salmonella, Pasteurella multocida and Streptococcus agalactiae were 100, 25, 50, 25 and 50 mg/mL respectively. The isolation of EA, EB and EC had no antibacterial effect on Escherichia coli and Pasteurella multocida. MBC of EA on Staphylococcus aureus and Salmonella were 2.5 and 5 mg/mL, and MBC of EB on Staphylococcus aureus, Salmonella and Streptococcus agalactiae were 1.25, 5 and 2.5 mg/mL respectively, while MBC of EC on Staphylococcus aureus was 2.5 mg/mL. Polygonum taibaishanense Kung.

PTKE对大肠埃希菌、金黄色葡萄球菌、沙门氏菌、多杀性巴氏杆菌和无乳链球菌的最低杀菌浓度分别为100，25，50，25和50 mg/mL；而PTKE的分离物EA、EB和EC对大肠埃希菌和巴氏杆菌无抑菌作用，EA对金黄色葡萄球菌、沙门氏菌的MBC分别为2.5和5.0 mg/mL，EB对金黄色葡萄球菌、沙门氏菌和无乳链球菌的MBC分别为1.25，5.0和2.5 mg/mL，EC对金黄色葡萄球菌的MBC为2.5 mg/mL。

Coli. Transmission electron microscopy was used to evaluate the cell morphology of both the normal and the treated E. coli. The observation with TEM suggested that silver nanoparticles lead to the formation of "pits" in cell wall of the bacteria, and silver nanoparticles could enter into periplasm through the pits and destroyed the cell membrane.

采用透射电镜观察了经纳米银粒子处理过的大肠杆菌细胞形态变化过程，结果显示纳米银粒子先在细胞壁上产生小的孔洞，通过这些孔洞进入周质空间，导致细胞膜成分渗漏和破坏细胞膜，进而进入细胞内部。

MethodsThe D. bacteria - E. coli shuttle expression vector pZT17 was constructed based on plasmids of pUE30, pGBM5 and pKatCAT. Then pZT17 with lux+ from Photinus pyralis was used to transform into D. grandis and E.coli. The recombinant strains were induced separately.

以质粒pUE30、pGBM5及pKatCAT为基础，构建抗辐射菌属一大肠杆菌间的穿梭载体，将groEL启动子和荧光素酶基因lux+插入到构建的穿梭载体中得到穿梭表达载体，并将该载体转化大肠杆菌诱导荧光素酶基因的表达。

METHODS:The gene of human ET1 was synthesized according to the preferential codons of E. coli, cloned into the EcoRI and SalI sites of vector pThioHisA. The recombinant plasmid pThioHisA-ET1 was constructed , sequenced and transformed into E.coli TOP10. Induced and expressed fusion protein were identified and analysed by 12% SDS-PAGE and densitometry analyses. After the elution, denature and renature, the fusion protein Thioredoxin-ET1 was obtained by ProBondTM chromatogragraphy. The purity of Thioredoxin-ET1 was detected by HPLC. Inoculate Thioredoxin-ET1 once per mouse every 2 weeks in 25, 50 and 100μg separately on 3 groups for 4 times. 10 days after last inoculation, we obtained venipuncture blood.

根据人ET1的多肽序列合成ET1基因，将其插入到pThioHisA的EcoRI和 SalI位点，重组质粒pThioHisA-ET1进行酶切鉴定及序列测定验证后转化TOP10，IPTG诱导的重组菌经SDS-PAGE检测融合蛋白Thioredoxin-ET1的表达量；表达的融合蛋白用ProBond亲合层析纯化并经HPLC鉴测其纯度；每只小鼠按25、50、100ug/次剂量的Thioredoxin-ET1每两周免疫一次，共4次，最后一次免疫10d后制备抗血清，经Western blot和ELISA检测证明Thioredoxin-ET1融合蛋白具有ET1免疫反应原性。

Coli. The outer membrane protein were extracted from 38 strains of avian E. coli which were isolated from the dead chickens of chicken breeding farms in 3 areas of Baoding, Qiuhuangdao and Beijing by using ultrasonic cleaving and N-Lauroyl Sarcosine Sodium, and OMP typing was done by SDS-PAGE.

方法］对分离自保定、秦皇岛和北京3个地区规模化养鸡场的病死鸡体内的38株大肠杆菌，采用超声波裂解和N-十二烷基肌胺酸钠进行外膜蛋白提取，利用SDS-PAGE进行Omp分型。

Listen,point and check your answers.

听，指出并且检查你的答案。

Warming needle is one of effective treatment methods for knee arthralgia aggravated by cold,and it is simple,safety,so it should be developed in clinical acupuncture and moxibustion extensively.

但以本院科针灸门诊在2005年1月—2006年6月期间共收治膝痛患者100余例，经过临床的诊断后，其中施以温针治疗的48例，疗效显著，报道如下。1临床资料本组病例48