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The research work involving the expression manner of doxA gene in S. lividans TK24 with plasmid pYG57 implied that the promoter PermE may control its expression constitutively, the time for maximum expression emerged from 48 to 60 h after inoculation and cultivation and the expression level was kept relatively stable, furthermore, the recombinant hydroxylase existed mostly in mycelium cell but little in broth.

而对Tm4（pYG57）菌株中doxA基因表达方式的研究表明， dd基因在 PermE启动子控制下可能是一种组成型控制表达，其表达量在接种后培养到 48－60 h左右最高并且维持相对稳定，并且表达的柔红霉素 Cl4羟化酶主要存在于菌丝体细胞内，很少分泌到细胞外。

The SDS-PAGE electrophoresis indicated that all engineered strains containing different expression plasmid could apparently express a 45 kD band specific for daunorubicin C-14 hydroxylase. The expression level of doxA gene was much higher when the doxA gene was under the control of promoter PermE. However, it was found that fd terminator had few effect on the expression of doxA gene.

SDS-PAGE蛋白电泳实验证明，上述五个菌株都能够表达大小约45 kD的柔红霉素C-14羟化酶蛋白，并发现以在PermE启动子控制下的doxA基因表达量相对较高，而终止子对＠中英文摘要 dd基因表达似乎没有影响。

Key words: Eimeria acervulina;3-1E gene;eukaryotic expression plasmid;immune protection

堆形艾美球虫；3-1E基因；真核表达质粒；免疫保护

Objective To construct recombinant plasmid of IgG2aVH region antisense RNA of systemic lupus erythematous. Methods Total RNA was isolated from spleen cell of BWF1 mice.

目的：构建系统性红斑狼疮鼠模型F1淋巴细胞IgG2aVH区反义RNA表达重组体，为SLE基因治疗提供基础。

In this paper, an escherichia coli-saccharopolyspora erythraea shuttle vector containing the erya promoter region was constructed using the enhanced green fluorescent protein gene as a reporter. the shuttle plasmid was transformed into sac.erythraea a226 and streptomyces lividans jt46 by peg mediation, respectively. fluorescence microscopy confirmed that the egfp was expressed in both strains.

本文克隆了erya基因的启动子perya，以绿色荧光蛋白基因为报告基因，构建了大肠埃希菌-糖多孢红霉菌穿梭型质粒。peg介导原生质体转化法将穿梭型质粒分别转入糖多孢红霉菌a226与变铅青链霉菌jt46，荧光显微镜检测发现，此启动子在两菌株中都具有功能。

As most Streptomyces promoters are invalid in S. erythraea, the promoter of erythromycin-resistant gene, ermE, and Streptomyces chromosomic integration site, attP, and apramycin-resistant gene, apr from plasmid pSET152 are utilized to construct the expression vector pZMW.

因大多数链霉菌质粒的启动子在糖多孢红霉菌中都不能发挥作用，故利用糖多孢红霉菌染色体上红霉素抗性基因PermE启动子作表达载体启动子，并利用pSET152质粒上链霉菌染色体整合位点和氨朴霉素抗性基因，构建了表达载体pZMW。

The multi-copy expression vector pZM is a shuttle plasmid which can replicate in Escherichia coli, Streptomyces and S. erythraea. It contains PermE promoter, fd terminator, multiple clone site, thiostrepton and ampicillin resistance genes, and ColE1 ori and pJV1 ori replicons which make the vector replicate in E.

游离型多拷贝表达载体pZM是可以在大肠杆菌、链霉菌和糖多孢红霉菌中扩增的穿梭型质粒，带有PermE启动子、fd终止子、多克隆位点、硫链丝菌肽和氨苄青霉素抗性基因以及在大肠杆菌和糖多孢红霉菌中复制的ColE1 ori和pJV1 ori复制子，载体大小9880bp。

In the genome of Klebsiella pneumoniae NTUH-K2044, we found nine fimbrial gene cluters including fim, mrk, and seven novel fimbrial operons, namely kpa, kpb, kpc, kpd, kpe, kpf, and kpg. These fimbrial gene clusters were cloned into the expression plasmid pET30b and then transformed to an afimbriated E.

在克雷白氏肺炎杆菌NTUH-K2044的基因体中，藉由HMMER找到了九套纤毛基因组，其中包含已知的第一型纤毛fim基因组和第三型纤毛mrk基因组，以及七套未曾被报导的纤毛基因组，分别被命名为kpa、kpb、kpc、kpd、kpe、kpf和kpg。

Suppression subtractive hybridization cDNA plasmid libraries were constructed between gastrula embryos and tail bud embryos in gynogenetic gibel carp.

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。

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。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！