质粒

It was not proved that the CpG of pUC19 could enhance the antigenicity of the recombinant plasmid pCMV-ME in this study.

用含CpG的pUC19质粒和重组质粒的共免疫与单独免疫重组质粒所诱导的抗体水平没有显著差别。

After removal of the episome, iPS cells completely free of vector and transgene sequences are derived that are similar to human embryonic stem cells in proliferative and developmental potential.

在质粒中的基因所表达的蛋白质会将细胞重新编程并使其成为iPS细胞。这些iPS细胞在接着的几轮细胞分裂中会开始失去这些质粒，这样研究人员就可以分离出不含质粒的细胞。

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。

Firstly, the pGL3-Control vector was reconstructed , the pGL3-Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN-β promoter gene was cloned into the pGL3-Enhancer vector and pGL-IP21, the Luciferase reporter plasmid with IFN-β promoter was OK. The availability of pGL-IP21 was verified by NDV ,the inductor of IFN-β, the Luciferase activity was assayed in cells transfected with pGL-IP21 by Luminometer.

首先将pGL3-Control载体进行了改构，除去了SV40启动子基因，获得了pGL3-Enhancer载体，将获得的IFN-β启动子基因连入载体中，构建了带有人IFN-β启动子基因的荧光素酶报告质粒IP-21，并且通过实验证明所构建的质粒在干扰素的诱导剂NDV的作用下能够表达荧光素酶活性，照度计检测荧光素酶活性增强，从而验证了所构建的重组质粒的有效性。

In order to develop a safe and effective immunoadjuvant to enhance the immunity and resistance of animals against infection, a novel CpG Oligodeoxynucleotides containing 11 CpG motifs was synthesized and inserted into the VR1012 plasmid, designated as VR1C. Then the recombinant VR1C was entrapped with Chitosan nanoparticles prepared by the method of ionic cross linkage, and employed to inject muscularly 3-weeks old Kunming mice; the blank VR1012 packed with CNP and saline were used to inoculate mice as the control groups. 28 days after inoculation, all mice were orally fed with 0.4ml 2x108CFU/ per mouse virulent hemorrhagic enteritis E. coli to challenge the resistance against infection.

为研制安全高效免疫调节剂增强动物免疫抗病能力，本实验设计合成含11个C pG基序的寡聚核苷酸，重组构建含CpG的VR1012质粒(VR1C)；制备壳聚糖纳米颗粒包裹重组质粒(CNP-VR1C)，肌注接种3周龄昆明小白鼠，设壳聚糖包裹空质粒和生理盐水对照组；接种后28天口服大肠杆菌攻毒观察小鼠天然免疫的变化和对强毒感染的抵抗力，Sandwich ELISA测定血清免疫球蛋白和白细胞介素含量。

The structure gene pelA from Thermotoga maritima MSB8 encoding pectate lyase was amplified and ligated into pHsh, resulting pHsh-pelA. Through structural optimization on pHsh-pelA, the ultimate plasmid, pHsh-pelC, which possessed the most appropriate structure and free energy of mRNA, was obtained.

将来源于极端嗜热菌属海栖热袍杆菌Thermotoga maritima MSB8的编码碱性果胶裂解酶的结构基因pelA与新型热激质粒pHsh连接，得到重组质粒pHsh-pelA，运用mRNA二级结构预测软件对pHsh-pelA的翻译起始区的二级结构进行优化，得到了具有最佳mRNA二级结构及自由能的质粒pHsh-pelC。

The function of hypothesis transport genes NMB0098 and NMB0097 were tried to study by way of gene knock out. The phototype of N. meningitidis strain BT878 was observed before and after gene NMB0098 was knocked out.

选用质粒pUC18为目的基因的载体，经过酶切、去磷酸化、连接、电穿孔法转导和抗生素筛选等步骤，分别将目的基因NMB0098和NMB0097与质粒相连，并在目的基因中插入标记基因—卡那霉素基因，构建盒式插入的重组质粒pNB0098-K和pNB0097-K。

The unoperated sides of the treated animals also served as controls. Six normal rats were treated as normal control group. Three different siRNA plasmid solution containing RC2-Ⅰ, MAFbx-Ⅱ, CON (50μl , 0.8μg/μl)was injected and transfected by electroporation as methods mentioned above, respectively. The changes of RC2 and MAFbx mRNA levels and RC2 protein levels after 3 days were determined by real-time quantitative PCR and Western blot, respectively. On postoperative 2, 3 and 4 weeks, the rate of wet muscle weight preservation, mean diameter of muscle fiber and mean cross-section area of muscle fiber and muscle protein content were checked and then compared between group CON and group RC2 or group MAFbx, respectively. The differences between groups were analyzed by one-way ANOVA. Ultrastructural changes of muscle fiber were observed at 2, 3, 4 weeks postoperation.Results GFP plasmid was efficiently deliverd into muscle by electroporation and robust GFP expression in muscle could be observed more than three weeks. Histology shows that injected plasmid DNA diffuses extensively in muscle tissue.

1、健康雌性SD大鼠18只，随机分为电穿孔组和非电穿孔组，每组9只，制作右下肢趾长伸肌失神经支配模型；EP组为将质粒pEGFP-N1溶液50μl(0.8μg／μl)注射入右趾长伸肌后，立即于两侧腱腹交接处给予电穿孔，电穿孔参数为：电场强度为200V／Cm，脉冲100μs，频率1Hz，施加10次脉冲；NEP组仅质粒pEGFP-N1溶液注射；转染后1、2、3周，荧光显微镜下观察趾长伸肌中GFP的表达情况，转染后1周行Western印迹检测趾长伸肌中GFP蛋白的表达情况，检测和优化体内转染效率。2、健康雌性SD大鼠78只，随机分为失神经对照组、RC2基因治疗组(RC2组)，MAFbx基因治疗组，每组24只，制作右下肢趾长伸肌失神经支配模型，余6只为正常组；分别将含CON、RC2、MAFbx基因的siRNA重组质粒注射入趾长伸肌，之后给予电穿孔，方法同上；治疗后3天实时定量PCR和Western印迹检测各组中RC2或MAFbx基因的mRNA和蛋白的表达变化，治疗后2、3、4周检测各组肌湿重维持率、肌细胞直径和肌细胞截面积，肌细胞超微结构变化以及肌纤维中蛋白含量变化。

BALB/c mice were immunized with plasmid TR421-hCGβ or mock DNA three times. Two weeks after immunization, spleen cells from the immunized mice were harvested, and then analyze the CTL activity against Sp2/0-ehCGβ cells. The splenocytes derived from the immunized mice were adoptively transferred to the normal mice, which were subsequently given injections i.

以TR421-hCGβ质粒实施基因免疫，免疫后检测小鼠脾细胞，特异性细胞毒活性；同期将脾细胞过继免疫给正常小鼠，以过继TR421-hCGβ质粒免疫小鼠脾细胞为实验组、过继TR421质粒免疫小鼠脾细胞为对照组，检测脾细胞杀伤效应。

Then the combinant plasmid was conducted into ACC-2 cell by electroporation. ACC-2 cells stably expressing CD was obtained by 10-day positive selection with 400 μg/mL G418. Total RNA was extracted and the expression of the CD gene in transfected ACC-2 cells was identified by RT-PCR. RESULTS: PCR yielded a fragment of l280bp and CD was verified by sequence analysis.

测序正确后，将其亚克隆到质粒表达载体pIRES中，构建以内部核糖体进入位点相连的CD基因的质粒表达载体pIRES-CD，采用电穿孔法，以质粒表达载体转染ACC-2细胞，用400μg/mL的G418筛选10d，获得稳定表达CD基因的ACC-2细胞系，提取该细胞的总RNA，用RT-PCR检测CD基因的表达。

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！