查询词典 cloning

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。

Result: Restriction enzyme analysis proved that cloning vector pEGFP-RECK was successfully constructed, and the sequencing results of it was in accord with expected. By transfecting the pEGFP-RECK vector into SHG44 cells, we could see the protein locate mainly in cell plasm.

结果：所得RECK基因融合蛋白表达载体序列与预期相符。pEGFP-RECK表达质粒转染SHG44细胞后，荧光信号主要集中在与细胞质有关的部位。

Adding ouiduet epithelial cell or granulose cell to CR1aa or TCM-199 co-culture, the cleavage rate, 4-6 cell developmental rate and 8-16 cell developmental aresignificantly increased (P＜0.05). We can conclude that: CR1aa and TCM-199 containing 5％FBS were the samewith somatic cell cloning in culture in vitro in Yanbian Yellow Cattle.

使用SPSS14.0软件对结果进行方差分析并多重比较后，综合考虑实验结果和方便操作时间与过程，得出以下结论：含5％FBS的CR1aa和TCM-199培养液适用于延边黄牛体细胞克隆胚胎的体外培养。

So cloning and marking the resistance gene of the downy mildew in cucumber are fundaments for disease resistance breeding of cucumber.

克隆和标记黄瓜霜霉病抗病基因，则是当前利用分子技术进行抗病育种工作的基础。

Objective To develop a new technique for efficient and rapid non-test tube cloning of the medicinal and energy-producing plant Jatropha curcas.

目的建立药用及能源植物小油桐非试管快繁技术。

In the 20th century,science has developed by leaps and bounds.although science is pushing human society forward ,its negative effect has resulted in many global problems ,the human are facing to environment pullution ,ecology unbalance ,agene technology and cloning man in the front of victory .which makes man confront the alternative of life or death.

20世纪，尤其是20世纪下半期，科学得到了更加迅速的发展，在科学推动人类社会进步的同时，它的负面效应也更突出并引发了全球问题，人类在胜利的面前又要面对全球性的环境污染、生态失衡、核威胁以及基因技术、&克隆人&等引发的冲击，使人类社会面临生死存亡的选择。

South Korean scientists say they have engineered four beagles that glow red using cloning techniques that could help develop cures for human diseases.

韩国科学家称他们运用克隆技术培育了四只发红光的猎狗，这项技术可以帮助治愈人类疾病。

When designing DNA cloning experiments, any possible biohazards associated with the creation of recombinations of genes must be contained, to minimize the potential risks to humans and to the environment.

在设计DNA克隆实验时，必须考虑到任何可能与基因的遗传因子重组有关的生物危害的产生，从而将对人类和环境的潜在危险减到最小程度。

Allen was a researcher at the Department of Neurobiology, The Babraham Institute, Cambridge, UK. Dr. Allen's research skills and techniques include DNA/RNA manipulation and purification/isolation, electrophoreses, PCR, cloning, all blottings, library screening, genetic mapping, design and construction of transgenes, artificial chromosome engineering, design and construction of vectors for gene targeting in ES cells, cell culture, histological techniques, mouse behavioural analysis, small animal surgery, and bioinformatic tools such as sequence databases and analysis tools.

从1996年到 2004年，Dr。 Allen 一直在英国剑桥Babraham 研究所神经生物学部门从事研究工作，包括DNA/RNA操纵、提纯/分离、电泳、PCR、克隆、印迹工作、文库筛选、遗传图绘制、转基因设计与构建、人工染色体工程、用于ES细胞基因打靶的载体设计与构建、细胞培养、切片技术、小鼠行为分析、小动物外科手术以及序列数据库和分析工具等生物信息工具的研究。

The monoclonal cells were selected using cloning rings. The expression of capsid protein was detected by indirect immunofluorescence and sandwich-ELISA. The empty capsids of FMDV were observed under electron microscope.

经间接免疫荧光和酶联免疫吸附测定（Enzyme-linked immunosorbent assay，ELISA）方法检测MDBK细胞中衣壳蛋白的表达，并在电镜下观察口蹄疫病毒空衣壳。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐