质粒

Through comparing the growth and exogenous gene expression of recombinant Anabaena sp.

比较无选择压力下连续传代的转基因鱼腥藻 712 0在不同培养基中的生长和外源基因表达，证实没有发生质粒部分缺失，但转基因鱼腥藻在无选择压力下会降低重组质粒拷贝数。

The successfully constructed new suicide plasmid pUC19-SacB can be used to construct a Brucella unmarked mutant.

构建了一个新的自杀质粒pUC19-SacB，该质粒能用于布鲁氏菌无痕缺失突变株的构建。

A conclusion can be drawn that tryptopanase expressed only by constructed plasmid during "catabolite repression".

在葡萄糖存在的条件下由于分解代谢产物阻遏，基因组上的色氨酸酶基因不表达，加入IPTG只表达质粒上的基因，如果在合适条件下测得色氨酸酶活性，则可以证明质粒上的基因能表达出有活性的色氨酸酶。

Methods 40 Wistar rats were divided into 2 groups(n=20): experiment group and control group.

40只雄性Wistar大鼠随机分为空质粒组和重组质粒组，每组20只。

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。

We composed the rhoa-gene of Wistar rat with the plasmid of pAdTrack-CMV to get the recombinant plasmid. On the basement, we can use the catholyte lipofetamine to transfected the neural stem cells , In the following, we observe the expression of the rho-gene and the geng how to act the motor neuron and NSCs.

本试验用克隆出的大鼠rhoa基因构建到质粒pAdTrack-CMV中，获得重组质粒pAd-rhoa，用阳离子脂质法介导转染到神经干细胞，观察目的基因的表达及其对运动神经元和神经干细胞的作用。

Plasmid DNA of pBILGC containing Chitinase and β-1, 3 - glucanase gene and fresh pollen of Papaver nudicaule were mixed in 11％ sucrose solution, then the pollen suspension were pollinated on stigmas of the Papaver nudicaule by artificial pollination.

以野罂粟开放花蕾为受体，以载有几丁质酶和β-1，3-葡聚糖酶基因双价植物表达载体pBLGC的质粒为供体，在11％浓度的蔗糖溶液中，加入花粉和含有目的基因的质粒DNA。得到药粉处理液。

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。

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！