原核

LP Recco PCR Cloning can efficiently and conveniently construct high quality prokaryotic expression vector of human NY-ESO-1 gene, which provide the expression vector to produce pET-15b-NY-ESO-1 fusion protein.

靶向克隆法可快速、高效地构建人NY-ESO-1基因的原核表达载体，为制备pET-15b-NY-ESO-1融合蛋白奠定了基础。

MethodUsing the canis MC4R DNA as template,the specific primers were designed. After PCR amplification,product was cloned into pGEX-4T-1 vector by LP Recco PCR cloning technique,the recombinant pGEX-4T-1-cMC4R was transferred to E.

方法以Beagle犬基因组DNA为模板，经PCR技术扩增目的片段，利用LP Recco PCR克隆技术将目的片段直接重组到原核表达载体pGEX-4T-1上，转化到E.coli DH5α，筛选阳性克隆，BamHI、XhoI酶切鉴定，DNA 测序检测插入序列的正确性。

Inclusion bodies are easily formed when recombinant proteins are expressed in E.coli system, and how to renature these inclusion bodies is now becoming the key problem in the genetic engineering.

E.coli作为目前应用最为广泛的原核表达系统，在异体表达蛋白质的过程中容易形成无活性的包涵体。

But 5-helix was apt to form inclusion body when expressed directly in prokaryotic cell and was difficult to renature, which causes inconvenience to future study.

但5-helix基因在原核细胞中直接表达时易形成包涵体，复性困难，给研究带来不便。

Two relevant sites of enzymatic digestion were added to the mTNF-α by PCR. The mTNF-α was linked to the 3'end of m/〓 in pGEX4T-1 vector. The prokaryotic expression vector pGEX4T-1m/〓-mTNF-α was constructed successfully. After induction and expression by IPTG, the expression of two kinds of fusion protein is 15% and 12% of total bacteria proteins respectively. The anti-HCC bifunctional antibodies m/〓-mTNF-α were identified by electrophoresis after the inclusion bodies were purified, denature, renature, re-purified, digested by thrombin and further purified.

采用PCR的方法在mTNF-α的两端加上所需要的酶切位点，将之连接在m／〓的3'端，构建原核表达载体pGEX4T-1 m／〓-mTNF-α，通过IPTG的诱导表达之后，两种融合蛋白的表达量分别占细菌总蛋白的15％、12％，表达产物经包涵体的纯化→变性→复性→纯化→凝血酶酶切→进一步纯化后，可以得到纯度为电泳纯的m／〓-mTNF-α抗肝癌双功能抗体。

A dominating antigenic region located at the amino-terminus of Hantaan virus nucleocapsid protein.These recombinant truncated proteins can be used to diagnose the infection of HFRSV serologically.

汉滩病毒核蛋白的氨基端含有一主要抗原决定簇区；原核表达的截短核蛋白在HFRS的血清学诊断中具有一定的价值。

Prokaryotic expression vector pET28b㈩/ TprK has been constructed successfully, and pET28b㈩/ TprK protein expressed to have set foundation for preparing prophylactic vaccine and developing serologically diagnostic kit for TP infection.

成功构建人pET28b㈩/TprK原核表达载体，并能高效表达TprK蛋白，为梅毒的预防性疫苗的制备以及进一步研究其血清学诊断试剂奠定了基础。

A complete bovine 18ku-bFGF gene was cloned by nested-PCR and subcloned into expression vector pET-28a.The recombinant plasmid pET-28a-bFGF was transformed into Escherichia coli BL21.At 25℃,recombinant protein was induced by 0.5mmol/L IPTG for 5 hours.Supernatant of cell lysate was purified using Ni-NTA method and the products was submitted to detect the bioactivity by Western-blotting,which indicated that the fusion protein contained recombinant bovine bFGF.Results showed that recombinant bovine bFGF was produced and solubly existed in the supernatant.NIH-3T3 fibroblasts were used to detect the bioactivity of the fusion protein.

采用巢式PCR方法克隆了牛18ku-bFGF基因完整的编码序列，并构建了原核表达载体pET-28a-bFGF，将其转化大肠杆菌BL21，在25℃低温条件下，用0.5mmol/L IPTG诱导表达5h，用Ni-NTA亲和纯化细胞裂解上清液，经Western-blotting检测，结果显示，在特定的诱导条件下，重组牛bFGF基因在大肠杆菌中获得了表达，并且主要以可溶性状态存在于细胞中。

Suum were reported to be suitable vaccine candidates for the control of Ascaris infection. Here we cloned As37 gene by reverse transcription-polymerase chain reaction using primers based on the pubmed cDNA sequence of As37 gene.

作者已克隆了猪蛔虫rAs37基因并进行了原核表达，并通过对重组融合蛋白As37进行免疫原性分析，显示其具有良好的免疫学活性。

After being induced to differentiate in vitro, cells with various morphologies showed β-gal activity, including nerve cells, neuroglial cells, epithelial cells. We also detected β-gal activity in a wide variety of tissue elements in solid tumors made by injecting the MC15 and MA34 cells into syngenic mice. Then we produced chimeric embryos by injecting the MC15 cells into Kunming blastocysts and recovering the embryos at various developmental stages.

SiCMVIE(Simian Cytomegalovirus Immediately Early)启动子是一个强启动子，本文首次将SCMVIE启动子应用到ES细胞途径上制作嵌合鼠，并通过lacZ基因在ES细胞和早期嵌合胚胎中的表达，表明该启动子是一个能在小鼠多数组织表达的启动子，这与1993年Peppel等人用原核显微注射法获得的转基因小鼠上所得的有关结果是基本上是一致的。

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