质粒

Fluorescence in situ hybridization was carried out to confirm the integration of HBV DNA into male pronucleus and its replication with cell division in embryonic development.(Reverse transcriptase polymerase chain reaction,RT-PCR) and immunofluoresence assay were performed to observe the expression of the HBV gene in two-cell stage.

材料与方法：成熟雄鼠麻醉后双侧睾丸注射经脂质体DOSPER包裹的HBV质粒，手术后雄鼠与超排雌鼠合笼交配，用荧光原位杂变(Fluorescence in situ hybridization，FISH)分别检测单细胞胚和二细胞胚间期核中HBV DNA的存在与复制，用逆转录聚含酶链反应(Reverse transcriptase-polymerase chain reaction，RT-PCR)和免疫荧光方法检测HBV基因在二细胞胚胎中的表达。

Objective To explore an experimental method of transfecting the marrow stromal stem cells with the reconstructed PGL3-transforming growth factor-β1 （TGF-β1） gene and to evaluate the feasibility of self-induction of MSCs to the chondrocytes in vitro so as to provide a scientific and experimental basis for a further ＂gene enhanced tissue engineering＂ research.

目的 探讨将编码细胞转化生长因子β1（transforming growth factorβ1，TGF—β1）的重组PGL3-TGF-β1质粒转染兔骨髓基质干细胞（marrow stromal stemceils，MSCs），体外通过自分泌、诱导作用向软骨细胞方向分化的可行性，为基因加强组织工程学修复软骨损伤提供体外实验依据。

METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction, and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.

从pEMSV-MyoD质粒上用聚合酶链反应法扩增出MyoD cDNA片段，再通过聚合酶链反应使MyoD基因加上CACC序列接头，经过定向拓扑克隆使目的基因连接到转移载体上，再通过LR酶促反应，将目的基因转移到腺病毒表达载体DNA上，获得MyoD基因重组的腺病毒DNA，用脂质体转染法转染HEK293A细胞，包装扩增出MyoD基因重组的腺病毒。

These transgenic cells were able to secrete hEGF protein to certain extent, and have the biological activities to enhance the growth rate of Hacat cells in vitro. Conclusion: Adult epidermal keratinocytes in vitro transfected with exogenetic hEGF cDNA can express and secrete active hEGF.

质粒pcDNA3.1-hEGF在脂质体介导下成功转染成人皮肤角质形成细胞，转基因细胞能分泌有生物活性的EGF；体外培养的成人皮肤角质形成细胞可见少量EGF分泌。

Decreased expression of MMPs by blocking the expression of Snail in ovarian carcinoma cells Objective: To investigate the influence of antiadhesive antibody (SHE78-7), targeted at the potent hemophilic cell adhesion molecule E-cadherin, on the motility and invasive ability of ovarian carcinoma cells that have stably tansfectant with anti-Snail plasmid.

第二部分反义Snail对裸鼠卵巢癌原位移植瘤转移的影响目的：观察转染反义Snail质粒的卵巢癌细胞在裸鼠原位移植瘤模型中生长及转移的情况，检测转移组织中Snail、E-钙粘蛋白、及基质金属蛋白酶的表达和分布。

Methods Femoral arteries isolated from the rabbits which had been fed with a high cholesterol diet and locally perfused with MM-LDL within the artery beforehand, were used as the models. Antisense MCP-1cDNA was transferred into the arterial wall by injecting recombinant LNCX-anti-MCP-1/liposomal complex in the femoral sheath and the periarterial tissue.

以实验性高脂血及轻度氧化LDL局部保留灌注的家兔股动脉为模型，采用DOTAP阳离子脂质体包裹LNCX-anti-MCP-1质粒，将反义MCP-1 cDNA定位导入家兔股动脉鞘及周围组织，以原位杂交、 Northern杂交及狭缝杂交法观察反义MCP-1基因的表达，应用扫描电镜观察单核细胞在动脉内膜粘附的情况。

Methods Ampicillin inhibitionof E. coli Top10 pcs+ was tested at first, and then b-lactamase activity in periplasm was examined.

然后再使用抗生素抗性分析、b-内酰胺酶的酶活测定以及Western blot杂交技术，分析质粒编码的b-内酰胺酶从细胞质到细胞间质的分泌情况。

By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells served as negative controls.

利用脂质体介导基因转染技术将含有SV40Tag的质粒pCMVSV40T/PUR转染原代胚胎前软骨干细胞，以未转染细胞作阴性对照。

In bioconv ertion experiment for spiramycin of No.5 transformant, the bioconversion product was analysed with TLC, bioautography, HPLC and mass spectrum.Results showed that the bioconvert product is propionyspiramycin. Propionyl acylase gene was l ocolized on 0.53 kb insert DNA fragment of recombinant pasmid pLJM95. Analysis result of DNA sequence showed that content of G+C is 68.2% for 0.53kb insert DNA fragment .

以No.5转化子对螺旋霉素进行生物转实验，其转化产物经薄层层析，生物显迹，高压液相色谱及质谱分析表明，与标准丙酰螺旋霉素一致，这说明 No.5 转化子具有将螺旋霉素生物转化为丙酰螺旋霉素的能力，丙酰化酶基因位于重组质粒，pIJM95 0.53kb 插入DNA片段上。

Object In this study we framed the refusion gene antibacterial peptide[ Cecropin A(l ~ 8)- Magainin (1 - 12),CA(1 ~ 8)-MA(l ~12),CAM] vetor and expressed it in the Escherichia.coli. Thenpurified the recombinded peptide in the Intein Mediated Purificationwith an Affinity Chitin-binding Tag-IMPACT system.

目的 构建重组基因抗菌肽[Cecropin A(1～8)-Magainin(1～12)，CA(1～8)-MA(1～12)，CAM]质粒载体并在大肠杆菌(Escherichia.coli)中表达，应用几丁质自切割分离纯化系统(InteinMediated Purification with an Affinity Chitin-binding Tag，IMPACT)分离纯化抗菌肽，并测定其抗菌活性。

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！