质粒体

Results:Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected CHO together with the pcDNA,which confered resistance to G418. The positive clones were tested by FIH.

结果：运用阳离子脂质体导入法将重组pALTER质粒与pcDNA3质粒共转染CHO，成功筛选出的阳性克隆经FIH证明突变CD59可在CHO细胞表达。

The synthesized cDNA is a continuous band less than 6. 0kb shown on x-film through α〓PdCTP which was inserted in the first strain of cDNA. The titer of the Ha-cDNA library was about 1. 5×10〓cfu/ml which could represent all protein-encoding sequences from Hz-AM1 cells; Namely, the cDNA library has representativity. After the successful construction of the Ha-cDNA library, the open-reading frame of vp39 gene of Heliothis armigera was then fused in frame with the Ga14-DNA-binding domain in the pGBT9 plasmid and the fusion protein Ga14-VP39 was expressed in yeast to be used as bait protein.

在成功构建棉铃虫细胞Hz-AM1cDNA文库的基础上，本文接着将棉铃虫核型多角体病毒衣壳蛋白VP39（HaNPV-VP39）基因克隆到酵母双杂交系统(yeasttwo-hybrid system)中的质粒pGBT9上。pGBT9上含有酵母菌转录激活因子GAL4 DNA结合域的编码区（GAL4-DBD），使克隆到质粒pGBT9上的HaNPV-vp39基因与GAL4-DBD基因融合，在酵母菌中表达DBD-vp39融合蛋白，该融合蛋白即为酵母双杂交系统中的诱饵蛋白。

Expression of M, NP, F and HN genes were detected and confirmed by the indirect immunofluorescence analysis. The syncytium formation was observed in the HeLa cells co-transfected with pCAGG-M, pCAGG-NP, pCAGG-F and pCAGG-HN recombinant plasmids at 48h post transfection.

纯化后的重组质粒通过脂质体单独转染HeLa细胞，经间接免疫荧光试验检测到了M、NP、F和HN蛋白的表达。pCAGG-M、 pCAGG-NP、pCAGG-F和pCAGG-HN重组质粒组合共转染HeLa细胞48h后，可以观察到明显的细胞融合现象。

Methods One hundred and eight Sprague-Dawley rats were divided into three troops: the troop of flaps, the troop of grafts and the troop of fat with thirty six of each.

将带有PCD-VEGF165的大肠杆菌接种到LB培养基中，通过碱分裂法制备PCD-VEGF165质粒，再用反蒸发法将质粒包埋于脂质体中。

An expression vector for one isoform of the mbC α1 subunit cDNA was constructed using the human cytomegalovirus promoter to direct expression and this expression vector was used in a novel transfection assay of primary cultures of bovine adrenal chromaffin cells.

为表达具有生物活性的钙离子通道，将mbC的一个异构体的蛋白编码区插入到由人的涎腺病毒启动子控制表达的质粒中。这个表达质粒

In this paper, an escherichia coli-saccharopolyspora erythraea shuttle vector containing the erya promoter region was constructed using the enhanced green fluorescent protein gene as a reporter. the shuttle plasmid was transformed into sac.erythraea a226 and streptomyces lividans jt46 by peg mediation, respectively. fluorescence microscopy confirmed that the egfp was expressed in both strains.

本文克隆了erya基因的启动子perya，以绿色荧光蛋白基因为报告基因，构建了大肠埃希菌-糖多孢红霉菌穿梭型质粒。peg介导原生质体转化法将穿梭型质粒分别转入糖多孢红霉菌a226与变铅青链霉菌jt46，荧光显微镜检测发现，此启动子在两菌株中都具有功能。

The successful transfection of perifollicular cells was quantified by elisa.

制备得到的质粒脂质体制剂能够将质粒转染进皮肤细胞并介导体内瞬时表达。

Comparison from the Germany isolate, phocine distemper virus type Ⅱ(PDV-2) and vaccine strain, the major difference is the long signal peptide domain while the mature protein exhibit high identity, and all of the 13 serine residues, four potential asparagine glycosylation sites and two hydrophobic regions supposed to affect the fusion function are completely identical. These data supports the view that F protein is more conservative than H protein in CDV.

三、犬瘟热病毒中国分离株囊膜糖蛋白基因免疫小鼠诱发的抗体应答经一系列步骤将CDV-YZ0101株的两囊膜糖蛋白基因定向导入真核表达载体pcDNA3.1中，DNA测序和酶切分析筛选阳性重组质粒克隆pcDNA-H和pcDNA-F，以重组质粒DNA和脂质体共转染COS-7细胞，用间接免疫荧光试验证实转染的COS-7细胞的胞浆中分别表达了CDV的H和F蛋白。

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。

Co-transformation of plasmid pGg-gfp and plasmid pCc1001 which harbors the complementary gene trp1 was conducted by the PEG-mediated protoplast transformation of the oidia of LT2, a tryptophan auxotrophic strain of Coprinus cinereus.

采用PEG介导法把表达质粒pGg-gfp与辅助质粒pCc1001（含有trp1基因）共转化进色氨酸营养缺陷型的灰盖鬼伞粉孢子的原生质体中。

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！