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The change of α1,2-FT activity in the cell line before and after the tranfection was confirmed by the determination of enzymatic activity. The changes of cell lipid and glucolipid, especially the change of type Ⅱ oligosaccharide, in the cell line before and after the transfection was determined by Thin-Layer Chromatography and TLC immunostaining method, respectively.

通过酶活性测定证明转染前后细胞系α1，2-FT活性的改变，采用薄层层析、薄层层析免疫染色方法测定转染前后细胞脂质及糖脂，特别是Ⅱ型寡糖的变化。

MSCs were cultured in a conditional medium in subculture including: dexamethasone, vitamin C, and β-glycerophosphate. At the same time, we constructed the eukaryotic expression vector containing VEGF165gene. VEGF165 gene was transfected into MSCs by means of LipofectAMINE?2000. The stably gene expressive cells were screened with G-418 for 14 days.

在脂质体介导下将构建成功的真核表达载体pcDNA3.1- VEGF165转染到诱导培养的MSCs，G-418筛选阳性细胞克隆，将阳性克隆细胞和未转染的MSCs共同接种于β-磷酸三钙上，体外培养7天后，植入原大鼠肌肉内。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

The CCAAT binding factor of brewer's yeast Saccharomyces cerevisiae has been shown previously to be a heteromeric complex containing HAP2, HAP3, HAP4 and HAP5 subunits. This factor can bind specifically to CCAAT-containing upstream sites within the promoters of numerous yeast genes, and then activate the transcription.

HAP转录因子(HAP2/HAP3/HAP4/HAP5)是存在于酿酒酵母中的一种异源多聚蛋白，它能与酵母中许多启动子上游的CCAAT盒专一性结合，以增强基因的转录。

The CCAAT binding factor of brewers yeast Saccharomyces cerevisiae has been shown previously to be a heteromeric complex containing HAP2, HAP3, HAP4 and HAP5 subunits. This factor can bind specifically to CCAAT-containing upstream sites within the promoters of numerous yeast genes, and then activate the transcription.

HAP转录因子(HAP2/HAP3/HAP4/HAP5)是存在于酿酒酵母中的一种异源多聚蛋白，它能与酵母中许多启动子上游的CCAAT盒专一性结合，以增强基因的转录。

These properties produce robust systems that limit the catastrophic consequences of transposon mobilization, which can result in the accumulation of deleterious mutations, changes in gene expression patterns, and conditions such as gonadal hypotrophy and sterility.

这些属性特征构成了一个限制由转座元件活动所造成的严重后果的系统，从而防止转座子侵袭所带来的突变积累，基因表达谱的改变，以及生殖腺发育不良和不育。

Because of lots of space voltage vectors are used impliedly to compose this algorithm, the THD of motor phase current, the ripple of stator flux and torque are lower, and then the drive system torque response can be more quick.

在此基础上分析多相感应电机矢量控制系统和直接转矩控制系统的稳态机械特性和动态转矩过渡过程，用来全面分析、比较这两种高性能控制方法的性能，研究二者的内在联系。

The confocal laser scanning microscopy revealed that the cyto-skeleton of MEL and MEL-M organized incompactly while the F-Actin organization of MEL-W was compact.

由激光共聚焦显微镜观察到，MEL和转突变型p53的MEL细胞骨架排列松散，而转野生型p53的MEL细胞骨架交联多且致密。

Results Established prostate cancer cell lines are eventually infectible by adenoviral vector. The ratio of vector/cell at which infection occurs depends on the specific cell line.

结果腺病毒载体可以转染所有已建立的前列腺癌细胞株，但是每种细胞株转染所要求的载体浓度以及暴露时间不同。

Non-viral vectors have important safety advantages over viral approaches, including their reduced pathogenicity and capacity for insertional mutagenesis, as well as their low cost and ease of production.

对于非病毒载体的研究主要集中在靶向性、转染方式和转染效率等领域，较少关注病毒载体介导目的基因定点整合和目的基因长期表达等方面的研究。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。