查询词典 recombinant

The omp17.3 gene of Brucella abortus was amplified by PCR and then the amplicon was cloned into the eukaryotic expression plasmid pcDNA3.1 to construct a recombinant plasmid pcDNA3.1-omp17.3. Then the recombinant plasmid pcDNA3.1-omp17.3 was transfected into COS7 cells, and the expressed OMP 17.3 was detected by Western-blotting.

采用PCR方法扩增了布鲁氏菌17.3ku外膜蛋白编码基因，并将该基因克隆至真核表达载体pcDNA3.1中，成功构建了真核表达质粒pcDNA3.1-omp17.3.pcDNA3.1-omp17.3转染COS-7细胞后，通过Western-blotting检测到了17.3ku蛋白的瞬时表达。

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。

We studied the effect of acetic acid on cell growth and recombinant porcine somatotropin for recombinant Escherichia coil.

在这里我们所探讨的是醋酸对重组大肠杆菌的菌体生长和重组蛋白质猪生长激素表现度的影响。

METHODS摘要: Molecular cloning techniques were used to construct the recombinant plasmid pLMP1p53mt containing mutant p53 gene and EB virus LMP1 gene. Linear recombinant plasmid pLMP1p53mt was transfected into mouse androgenesis of germ cells by microinjection and then survival germ cells treated were planted into fallopian tubes of artificial pregnant female mice.

方法摘要：通过分子克隆技术构建含突变型p53基因和EB病毒LMP1基因的真核表达载体pLMP1p53mt；采用显微注射法将线性化的表达载体注射至小鼠受精卵的雄性原核中，然后将注射存活的受精卵植入假孕母鼠的输卵管；取其产3 wk龄子代鼠进行PCR初选，再通过Southern杂交进一步确证；利用HE染色法观察4 mo龄转基因小鼠不同组织病理变化。

Recombinant plasmid pFf-22-Trp operon was successfully constructed, and both the anthranilate synthase and tryptophan synthase activities of expressed product increased. It laid a foundation of construction of recombinant E. coli strain for production of tryptophan in a large scale.

已成功构建了重组表达质粒pET-22b-Trp operon，邻氨基苯甲酸合成酶和色氨酸合成酶的活性在大肠杆菌中得到了提高，为高产色氨酸基因工程菌的构建奠定了基础。

Methods A recombinant antisense GFAP retrovirus was developed. After packing and concentrating, the suspension of this retrovirus was injected into the sites of injured brain stab by a microinjector. The effect of recombinant antisense GFAP retrovirus on formation of glial cicatrix in the injured sites was studied through immunohischemistry and morphological observation.

构建反义GFAP逆转录病毒表达载体，经过病毒包装后，将获得的病毒上清液浓缩并通过微量注射的方法，引入大鼠脑穿刺损伤灶内，采用免疫组化及形态学观察的方法，研究重组反义GFAP逆转录病毒对脑损伤后胶质瘢痕形成的影响。

Recombinant ptotein made up 39.2%(pET-20b-bglA) and 33.5%(pET-28a-bglA) of the total proteins in the intracellular fraction, the solubility proportion of the enzyme up to 32.8%(pET-20b-bglA) and 40.1%(pET-28a-bglA), the activities of the enzyme were 66.8 (pET-20b-bglA) and 71.2 U/mg (pET-28a-bglA). These results showed that E. coli BL21-CodonPlus (DE3)-RIL with argU tRNA, ileY tRNA and leuW tRNA genes helped to improve expression of the enzyme through enhanced identification of the rare codons AGA/AGG, AUA and CUA. Further optimized the conditions for inducing, the solubility proportion of the enzyme was 70.6% at 16 °C and 1.2 times higher than 37 °C. The solubility proportion of the enzyme increased from 12.3% to 32.8% when IPTG concentrations dropped from 1000 μM to 25 μM. The recombinant enzyme was purified by heat treatment, DEAE Sepharose anion-exchange chromatography and TOYOPEARL HW-55F.

maritima MSB8 的-葡萄糖苷酶基因 bglA克隆至表达载体 pET-20b 和 pET-28a，构建重组质粒 pET-20b-bglA 和 pET-28a-bglA，然后转化不同大肠杆菌 E.coli 宿主，Tm-SIGlA 在 E.coli BL21-CodonPlus(DE3)-RIL 中获得高效表达，重组蛋白的表达量为 33.5%(pET-28a-bglA)和 39.2%(pET-20b-bglA)，可溶性比例为 32.8%(pET-20b-bglA)和 40.1%(pET-28a-bglA)，比酶活达 66.8 (pET-20b-bglA)和 71.2 U/mg (pET-28a-bglA)，这些结果表明，E.coli BL21-CodonPlus(DE3)-RIL 宿主带有的 argU tRNA、ileY tRNA 和 leuW tRNA 基因，分别增强对稀有密码子 AGA/AGG、AUA 和 CUA 的识别，有助于提高该酶在 E.coli 中的表达；进一步优化诱导条件，重组 E.coli BL21-CodonPlus(DE3)-RIL/pET-20b-bglA 在 37 ℃下诱导培养，IPTG 浓度由 1000 μM 降至 25 μM，目的蛋白可溶性表达由 12.3%增至 32.8%，提高 1.7 倍，16 ℃下低温诱导实现目的蛋白 70.6%的可溶性表达，较 37 ℃下诱导培养提高 1.2 倍。

A recombinant protein of Bla g 2 has been obtained, which is soluble in the supernatant and therefore can avoid a process of denaturalization and renaturation of the recombinant.

获得了真核表达的德国小蠊主要变应原重组蛋白Bla g 2，该蛋白以可溶性蛋白的形式分泌到培养液中，避免了原核表达的变复性过程。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

I'm strongly against the death penalty — it's an eye for an eye.

我不赞成死刑——这是以牙还牙的报复行为。

And to get you the support you need, we're enlisting all elements of our national power: our diplomacy and development, our economic might and our moral suasion, so that you and the rest of our military do not bear the burden of our security alone.

并给你们所须的支援，我们正徵召国家所有各种的力量：我们的外交及发展，我们的经济力量与道德劝说，所以你们与其他军人不须要孤独地负起国家安全的责任。