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Lamin A gene cDNA lacking of 150 basyls at the end of the eleventh extron was cloned into pcDNA3.1 carrier, and the recombinant plasmid pcDNA3.1-mutant LMNA cDNA was injected into the male pronucleus of mouse zygotes, then the zytotes was replanted into the oviducts of pseudopregnant mice.

实验方法:将缺失了第11外显子末端150个碱基的LMNA基因cDNA克隆到pcDNA3.1载体,应用原核显微注射方法将线性化的pcDNA3.1-mutant LMNA注射到小鼠受精卵的雄原核,再将注射后的受精卵移植至假孕鼠的输卵管。

Recombinant expression vector k14/c-myc-paav was constructed bearing the mouse c-Myc cDNA derived by the skin specific human keratin 14 promoter, then the linear vector was microinjected into the two pronucleus of the C57BL/6 fertilized eggs to produce transgenic mice.256 microinjected eggs were transferred, 6 of the foster mothers were pregnant and 44 adult mice were produced, two of them were positive transgenic mice confirmed by PCR and Southern blot analysis, transgenic rate was about 4%.

以CMV-PAAV载体为基础载体,以皮肤特异人K14(人角蛋白14)基因启动子替换原质粒中的CMV启动子后,在其多克隆位点区依次插入鼠c-Myc基因的cDNA序列和gfp基因的编码序列,构建了高效表达c-Myc基因的真核表达载体。通过双原核显微注射技术,将线性化的载体DNA注入到C57BL/6小鼠受精卵的两个原核中,然后将注射后状态良好的胚胎移植到代孕鼠的输卵管中,共移植了256枚受精卵,6只受体怀孕,最终获得44只成年小鼠。

PDGF-BB is the most effective subtype in the PDGF family on the metabolism of osseous tissue or the treatment of wound healing, and its clinical application was approved firstly among the growth factors although it was used to treat the decubital ulcer of diabetes.

本研究通过分子克隆方法获取人血小板衍生生长因子B链成熟肽基因,构建原核表达载体;利用大肠杆菌体系原核表达人血小板衍生生长因子B链前体蛋白,并纯化、复性进而获得具有功能活性的PDGF-BB;并对获得的重组rhPDGF-BB蛋白进行体外功能活性鉴定,同时,初步探讨PDGF-BB在毕赤酵母中的表达,为寻找更优化的表达纯化方法提供实验学依据。

To investigate the expression, location and function, long distance PCR was used to expand the targets containing open reading frame. The expressive vectors of prokaryotic and eukaryotic cells were constructed after product purification and connecting with vectors. The prokayotic vectors were induced to express the protein by IPTG and the protein was seen by denaturalization PAG electrophoresis and dyeing. The eukayotic expressive vectors were transfected into culture cells and the protein was found in the nulceolus under the observation of the confocal microscope. The transfected cells were chosen by the geneticin (G418). The cell cyele was examined by flow cytometer and the cell numbers were reduced obviously in the stage of G〓-G〓 and G〓-M, but increased distinguishably in the S stage.

为研究磷酸化应激诱导蛋白的表达、定位和功能,采用了长距离聚合酶链反应扩增含有开放阅读框架的靶片段,经产物纯化、与载体连接,分别构建了STIP1基因的原核表达载体和真核表达载体;IPTG诱导原核表达载体,变性聚丙酰胺凝胶电泳与染色后,可见相应大小的蛋白质表达;STIP1基因的真核表达载体转染细胞系后,共聚焦显微镜下观察STIP1蛋白主要分布于细胞核内;用药物筛选STIP1基因转染的细胞后,流式细胞仪检测细胞周期,可见G〓-G〓期和G〓-M期的细胞明显降低,S期细胞明显增多。

In order to develop antibacterial peptide biologics for prevention and control of endometritis of dairy cow,a pair of primers was designed and synthesized according to the sequence of bovine antibacterial peptide gene published in GenBank.The total RNA was purified from the dairy cow endometrial cells with the Total RNA Isolation Kit.

为研发防治乳牛子宫内膜炎的抗菌肽生物制剂,根据已发表的牛抗菌肽基因序列设计了1对引物,用试剂盒提取乳牛新鲜子宫内膜总RNA,经RT-PCR扩增,将克隆的抗菌肽BNBD5基因全长编码区的cDNA连接到原核表达载体pET30a中,经IPTG诱导原核表达,通过Western-blot检测融合蛋白BNBD5的表达水平。

The main objective of the dissertation was to improve and modify the experimental procedures of mouse nuclear transfer and pronuclear transplantation, respectively, and try to establish new possible methods for mouse cloning and pronuclear transplantation.

本论文的主要目的是改进和修改小鼠核移植和原核置换的实验程序,试图建立新的小鼠核移植和原核置换的可行方法。

The recombinant protein were purled from prokaryotic bacteria hadactivity of binding to IBV in ELISA and Western blotting test, which could be blocked byantiserum against gAPN.

将原核表达的gAPN蛋白进行的ELISA及Westem blotting试验证实原核表达的gAPN蛋白在体外可以与IBV结合,而且这种结合可以被针对gAPN的抗体阻断。

There are many documents indicated many kinds of pathogenic bacteriums have the ability to lead latent infection, maybe which is related with the genome conservatived TA system of prokaryote.The TA system is made up of toxin and antitoxin and is concerned to be quality control system of prokaryote.

研究结果提示,很多不同种属的病原菌都具有形成潜在感染的能力,可能与原核生物染色体基因组上保守的TA系统有关,其主要由毒素和抗毒素位点组成,被认为是原核生物基因表达的质量控制系统。

Complicated than procaryotes. These cells, exemplified by the yeast in Figure 4.1-3, have a membrane surrounding the cell which is similar in structure to the procaryote membrane.

真核细胞,具有真正的细胞核,其结构要比原核生物复杂的多,以图4.1-3中所示的酵母菌为例,和原核细胞一样真核细胞也具有一层细胞膜。

In most species they are composed of roughly equal amounts of protein and ribosomal RNA , the ribosome consists of two unequally sized rounded subunits arranged on top of each other like a cottage loaf, eukaryotic cells have larger ribosomes than prokaryotic cells but the ribosomes in mitochondria and chloroplasts are about the same size as prokaryotic ribosomes.

在大多数的物种中,蛋白质的量和 rRNA 量是大体相等的。核糖体包括两个大小不同的亚单位,上部结合形成小屋状结构。真核生物细胞中的核糖体要比原核细胞的核糖体大,但是线粒体和叶绿体中的核糖体与原核细胞的核糖体大小相当。

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