质粒

Suppression subtractive hybridization cDNA plasmid libraries were constructed between gastrula embryos and tail bud embryos in gynogenetic gibel carp.739 and 816 PCR positive clones were respectively selected to perform dot blot,and 72 dot blot positive clones and 98 dot blot positive clones were obtained from the SSH plasmid libraries specific for gastrula embryos and tail bud embryos.

构建了雌核发育银鲫原肠期胚胎和尾芽期胚胎间的抑制性差减杂交cDNA质粒文库。对原肠期739个和尾芽期816个PCR阳性克隆进行斑点杂交，得到72个原肠期和98个尾芽期斑点杂交阳性克隆。

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003～2007年临床分离的220株Pa，对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况，同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration，MIC)，筛选出对亚胺培南耐药的铜绿假单胞菌，并分析其对其它抗生素的药物敏感率；采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株，采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因，采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因，用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况；同时对产AmpC酶的Pa(25株，含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

In the research, one kind of guzmania species named ostara was employed to be as material, and the full-length cDNA plasmid library of floral organ was constructed successfully by using SMART technology. The library had 3×10^6 original titer and more than 1 kb insert fragments. After 5'EST sequencing from 2004 positive clone chosen at random and clustering analyse, 1 758 high-quality sequences and 1 365 unigenes including 175 contigs and 1 195 singlets were obtained. These unigenes had 1 283 valid ORFs. COG analysis showed that those proteins coded by EST sequences were divided into 22 classes. Through blast analysis, full length or part cDNA of some genes controlling flower color, development of floral organs, florescence regulation and other breeding value genes were obtained.

本研究以擎天凤梨属品种Ostara为材料，采用SMART技术成功构建了擎天凤梨花器官的质粒型全长cDNA文库，初始文库滴度为3×10^6，插入片段平均长度大于1kb；随机挑取2004个阳性克隆进行5'EST测序，获得高质量序列1758条，经拼接获得1365条单基因簇，其中跨叠群175个和单条序列1195个；经ORF寻找共获得有效ORF1283条；经COG分析EST序列编码的蛋白质被分为22类；经Blast分析后，获得一批花色、花器官发育、花期调控以及其它育种价值基因（包括cDNA全长或片段）。

Results The expression plasmid of pVAX1 IL-18 HN was contructed successfully confirmed by enzyme digestion identification. The expressions of IL-18 and HN chimeric genes were confirmed, and the expressed HN protein had higher hemagglutinative titer.

结果：酶切鉴定证实正确地构建了pVAX1 IL-18 HN嵌合表达质粒，Western blotting和血凝试验检测结果表明，嵌合后的IL-18和HN基因均能够表达，且表达的HN蛋白具有较高的血凝活性。

Western blot analysis indicated that all of the fusion proteins were expressed in AO3_ 1 cells, in which multiple copies of the lac operator were integrated to produce a heterochromatic region of the genome.

Western blot检测表明，这些重组质粒分别转染AO3_1细胞后均表达了相应的融合蛋白。

Positive rate was relatively high, and the foreign gene could also maintain in testicular tissue for a long time, it could still be detected in the tissue in 7 months after being injected. However. Deferent results could be gained when the detection were made with different primers on different sites of the recombinant plasmid. This could be explained by transgene lost when the foreign gene recombinated and integrated in heterogenetic cell, or transgene degradated by cell nucleases.

然而，我们也看到，在后代检测中，利用不同的引物扩增重组质粒不同部位时，检测结果存在差异，转基因阳性率不同，这种差异说明当外源基因进入到异源细胞内，在重组整合时发生了外源基因丢失的现象，并由此导致外源基因的整合率下降，也可能是受细胞内核酸酶的作用，使得外源基因遭到降解。

This article is to make use of the monolayer membranes and liposome to study that interaction of hypericin with model membrane to understand the influence of hypericin on the biomembrane.

本文是利用分子单层膜与微脂质粒系统探讨金丝桃素与拟生物膜之间的交互作用，来了解金丝桃素对生物膜的影响。

Traditional separation method: Several years ago, as a result of the equipment condition limit, the material particle DNA separation with the CsCl balanced equilibrium density centrifuge process, the idiomorphic become generally the gradient.

传统的分离方法：数年前，由于受设备条件限制，质粒DNA的分离一般用CsCl平衡等密度离心法，自形成梯度。

In conclusion, a prokaryotic plasmid, encoding the fusion gene of single-chain variable fragment with MCP-3 and expressing idiotype protein vaccination against B cell lymphoma, was constructed correctly.

本研究成功构建鼠源scFv片段与趋化因子MCP-3融合的独特型B细胞淋巴瘤疫苗表达质粒pGLo/scFv- MCP-3，并实现融合蛋白的初步表达。

The recombinant plasmids were analyzed by sequencing and the one containing the correct fragment was transfected into K562 cells. The condition of idiotypic protein expression was tested by Western-Blot.

将经测序鉴定正确的重组质粒转染K562细胞，Western-Blot等方法检测其蛋白表达的情况。

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