腺病毒

The recombinant adenovirus was then large amplified and purified with Viraprep purification column and desalted.

大规模扩增重组腺病毒并用Vira-prep纯化柱纯化。

Objective To improve the construction of adenovirus expression vector harbouring widetype tumor suppressor gene PTEN.

目的 为改进构建野生型抑癌基因PTEN重组腺病毒表达载体的方法。

Methods Bladder cancer cell line 253J and human fibroblast cell line MRC-5 were transfected by the recombinant adenovirus of different multiplicities of infection,and the infection rate was measured by observing the expression of enhanced ...

利用不同感染复数的重组腺病毒携带增强型绿色荧光蛋白基因感染膀胱癌细胞253J和人成纤维细胞MRC 5，荧光显微镜下观察其感染效率。

Results UCMSCs could be effectively infected by adenovirus containing GFP gene in vitro, the transfection efficiency has the dose-effect relationship with multiplicities of infection.

结果 腺病毒介导的GFP基因对于UCMSCs具有较高的转染效率，转染效率与MOI值具有量效关系，当MOI为100倍时，转染效率达95%。

The humoral response to hepatitis B surface antigen was mildly induced and could be enhanced by reinjecting a regular dose of HBsAg antigen vaccine.

初步结果显示腺病毒载体是一种高效、安全的载体系统。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

The second part of this study: the constrction of the recombinant adenovirus.

论文的第二部分：含内皮抑素基因的重组腺病毒的构建。

Objective To evaluate the inhibitory effect of recombinant adenovirus rAd-MK-TNFα on cell growth of breast cancer in vitro.

目的 初步观察重组腺病毒rAd-MK-TNFα对乳腺癌细胞的抑制作用。

Aim of the study is to construct urothelium-specific recombinant adenovirus and investigate its inhibition in bladder cancer cell.

本研究是构建膀胱上皮特异性重组腺病毒，探讨其在膀胱癌细胞中的抑制作用。

The infection efficiency of adenovirus is determined by the levels of CAR and integrin on cancer cells.

肿瘤细胞表面CAR和整合素的表达水平决定了腺病毒对肿瘤细胞的感染效率。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。