大肠杆菌

The unfused human interferon aD expressed by pBV867 was confirmed by SDS-PAGE,activity assay and N terminal sequencing.Results suggested that E.Coli is able to process the signal sequence of human interferon αD.

发现&翻译联结&方式起始的基因表达水平比一般ATG起始方式高约10倍；证明大肠杆菌能够识别并加工人αD型干扰素信号多肽。

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

It was found that the unstimulated human peripheral blood leukocytes only expressed a low level of MEFV mRNA.

RT-PCR扩增后的MEFV基因片段成功导入质粒载体，并且可以在大肠杆菌中培养生长。

In this study we constructed the recombinant pcDNA-HA-rpb in E.coli cells and we expressed the three polymerase II subunits; Rpb5, Rpb6 and Rpb7 as HA-tagged polypeptides by transfection in 293T mammalian cells, as well as untagged nuclearβ-actin.

在这项研究中，我们在大肠杆菌中构建了pcDNA-HA-rpb重组质粒，表达三个聚合酶II亚基： Rpb5 ， Rpb6和Rpb7，通过转染哺乳动物细胞293T，从而表达带HA标签的多肽，同时转染不带标签的β-actin。

Diarrhoea-associated haemolytic uraemic syndrome is most commonly caused by E.

腹泻相关性溶血尿毒综合徵最常见的致病菌是 O157：H7 大肠杆菌。

Methods: The bacteria inhibiting experiment in vitro was used to observe the bacteria inhibiting effects of usnic acid material and SMEDDS. The cytotoxic action on peritoneal macrophage and the mouse peritoneal macrophage releasing TNF-α induced by thermal inactivation bacillus coli of usnic acid material and SMEDDS were determined by MTT.

采用体外抑菌实验观察松萝酸原料及自微乳的抑菌效果；采用MTT法考察松萝酸原料及自微乳制剂对腹腔巨噬细胞的细胞毒作用，及对热灭活大肠杆菌诱导小鼠腹腔巨噬细胞释放TNF-α的影响。

The anti-inflammation test showed that within the concentration of 1-100 μg/ml both usnic acid material and SMEDDS had no cytotoxic action. The usnic acid material and SMEDDS with different dosage could bring the levels of the mouse peritoneal macrophage releasing TNF-α induced by thermal inactivation bacillus coli down, and it showed a dose-dependent relationship.

抗炎实验表明，松萝酸原料及自微乳在1~100μg/ml质量浓度范围内对小鼠腹腔巨噬细胞无显著细胞毒作用；不同质量浓度的原料药和自微乳制剂均可使热灭活大肠杆菌诱导的TNF-α水平有不同程度的下降，并呈一定的量-效关系。

In order to test the antibodies to E6 and E7 induced by the recombinant vaccina virus, the fusion proteins of GST and E6 or E7 were expressed in E. coli using pGEX-2T vector. The expressed proteins were confirmed by Western-blot and purified by glutathione sepharose CL-4B affinity chromatography. The purified proteins were used to detect the anti-E6 and-E7 by ELISA.

为便于检测重组痘苗病毒所诱发的E6和E7抗体，本文利用原核表达载体pGEX-2T，在大肠杆菌中分别高效表达了GST与E6或E7的融合蛋白，并利用纯化的融合蛋白成功地建立了E6和E7抗体的ELISA检测方法。

Furthermore. the bilirubin oxsidase activity was detected in the supernatant of cell lysates from IPTG induced culture of E. coli containing pEI-3a/BOX These results demonstrated that bilirubin oxidase cDNA from Myrothecium Verrucaria has been successfully expressed in E. coli.

这些结果表明，疣疱漆斑菌胆红素氧化酶cDNA首次成功地在大肠杆菌中获得了表达，所表达的可溶性产物具有与天然蛋白质基本相同的分子量、相似的免疫性和酶催化活性

Enterohemorrhagic Escherichia coli strains are zoonotic pathogens responsible for a range of severe human disease.

肠产毒性大肠杆菌菌株是动物传染性病原体并造成人类一系列严重疾病。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。