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Construction of eukaryotic expression plasmid of VEGF165 gene and its expression in myocardial cells ［J］.

VEGF165表达载体的构建及其对人胃癌生长的作用［J］。

The construction of the recombinant plasmid carrying shRNA to HPV6bL1 lays the basis for the study of its inhibitive effect on cauliflower excrescence.

经测序，测序结果均与目的序列相同，均符合设计要求。结论 HPV6bL1 shRNA质粒重组体的成功构建为研究尖锐湿疣基因靶向抗体打下基础。

After 2 days incubation, harvested exoenzyme RSDA from Bacillus subtilis DB430 with the recombinant plasmid pHR showed the best enzyme activity, around 6.5 U/mL. Crude RSDA is added into 20% ungelatinized Tainon 57 sweet potato powder for hydrolyzation and achieve the highest glucose yield 36 g/L.

Bacillus subtilis DB430 菌株在培养2天后可以达到大约 6.5 U/mL左右的最高酵素活性，经离心将粗酵素液直接加入 20%未经糊化的台农57号甘薯生粉中，最多可以产生36 g/L的葡萄糖。

Three RNAi target sites (named as Abil1, Abil2, Abil3) targeting the E3B1 (NCBI: NM-024397), an identified target sites and positive control GAPDH-A were selected. The pGenesil-1 eukaryotic expression vectors with the neoR mark and GFP green fluorescent mark were selected to construct E3B1 RNAi plasmid. The effect of RNAi targeting various sites on E3B1 genes expression was evaluated by testing the transfection efficiency with fluorescence microscope and western blot. The most effective siRNA in inhibiting E3B1 genes were screened. The most effective siRNA in inhibiting E3B1 genes screened and used to transfect the neuronal cells. After that, the inhibitor of axon growth was added and the axon growth was observed. Result: The siRNA expressing plasmids targeting the E3B1 were constructed successfully.

选择针对E3B1(NCBI： NM-024397)RNAi的3个靶位点Abil1、Abil2、Abil3和1个不针对任何mRNA的RNAi靶位点以及甘油醛－3-磷酸脱氢酶，用带有neoR选择标志和GFP绿色荧光标志的真核表达载体pGenesil-1构建E3B1 RNAi质粒，分别转染培养的神经元，在荧光显微镜下观测转染率，经G418筛选得到单一的转染细胞，并用Western blot法检测各转染组神经元E3B1蛋白的表达情况，选出具有最佳抑制效应的siRNA转染神经元，加入轴突生长抑制物，观测轴突生长情况。

The 528 bp fragment of flic gene was successfully cloned into vector, the pGEX-flic recombinant plasmid was expressed and induced by IPTG and Western-blot method showed good antigenicity of the recombinant protein, and there were flexible regions such as coil region and turn region in flic protein.

获得的目的基因片段长度为528 bp，测序表明其序列与GenBank公布的一致；软件预测结果显示，该蛋白具有多个明显的二级结构成分及跨膜区；SDS-PAGE检测表明，在约50 ku处有一特异性条带；Western-blot检测表明，表达蛋白具有良好的抗原活性。

PART FOUR EFFECTS OF MIL-4RA EXPRESSION VECTOR ON ASTHMATIC AIRWAY INFLAMMATION AND TH1/TH2 CELL DISFUNCTION THROUGH INTRAPERITONEAL ADMINISTRATION In the study of part three, we treated the mouse asthmatic model by intratracheal administration with mIL-4RA expression plasmid and observed obvious therapeutic effects. But intratracheal administration belongs to one of the insult therapies.

第四部分：腹腔注射mIL-4RA重组载体对哮喘小鼠气道炎症及Th失衡的干预作用前一部分研究中我们通过气道局部干预途径，即气管内直接滴注的方式，观察到mIL-4RA真核表达质粒对哮喘小鼠具有很好的治疗作用，但气管内滴注毕竟属有创性治疗，存在需要实验条件高，有较大的风险性，不易反复进行等缺点，这势必大大限制其在动物实验及进一步临床研究中的实际应用价值。

Lividans. Using pIJ903 bearing tsr gene as a vector, the two furthermost fragments of the cluster, 4. 0kb and 1. 5kb in size respectively, were inserted into it. To facilitate the detection of gene replacement, apr gene was placed in the middle of the two inserts. OriT from E coli plasmid RK2 was also incoporated into the vector, therefore, pIJ903 derivatives can be mobilized from E.

在具有硫链丝菌素抗性基因的pIJ903的单—BamHI位点，同向插入了FR-008生物合成基因簇两个最远端的4.0kb和1.5kb片段，并在二者之间插入了可在链霉菌FR-008和变铅青链霉菌中表达的阿泊拉霉素抗性基因，同时也插入了有助于将质粒引入链霉菌的oriT片段，从而构建出了置换整个基因簇的基因置换质粒pHZ691。

To evaluate the safety of the DNA vaccine pVAX1/F1-V against plague, the purity of plasmid DNA pVAX1/F1-V was detected by SDS-PAGE and gelose electrophoresis method; pasmid DNA was detected by PCR in tissues of BALB/c mice immunized with the plasmids intramuscular injection.

为了研究鼠疫DNA疫苗pVAX1/F1-V质粒的安全性，试验运用SDS-PAGE电泳和琼脂糖电泳等方法检测了鼠疫DNA疫苗pVAX1/F1-V质粒的纯度，并以小鼠为动物模型进行了质粒pVAX1/F1-V的急性毒性和长期毒性试验，用PCR方法检测外源基因在小鼠组织中的分布情况，用ELISA、ELISPOT方法检测了其自身的免疫反应。

Agrobacterium method was used to transform the plasmid into Gerbera hybrida.

用农杆菌介导的方法将该质粒转化到非洲菊中。

Inthe group injected with plasmid in 10% glyceryl,increase of GH was significantly deferentfrom control in 5d after injection.

其中在甘油组(质粒注射液中含10％的甘油)注射后第5天与对照组经t检验分析相差显著(P＜0.05)。

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！