筛选

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 2nd pace, right attack this IP safe strategy, in attribute dialog box, use the hook take out that adds guide left, click next add pushbutton to add new regulation, shoot a dialog box of new regular property subsequently, click on the picture add pushbutton, play an IP to choose implement list window; is in list, above all use the hook take out that adds guide left, the accretion that clicks right again next pushbutton adds new filtration implement.

第二步，右击该IP安全策略，在&属性&对话框中，把&使用添加向导&左边的钩去掉，然后单击&添加&按钮添加新的规则，随后弹出&新规则属性&对话框，在画面上点击&添加&按钮，弹出IP筛选器列表窗口；在列表中，首先把&使用添加向导&左边的钩去掉，然后再点击右边的&添加&按钮添加新的筛选器。

The a -Gal remarkably positive staining was appeared in membrane and plasm on ES cell, but nuclear is negative. It indicates that we must pay attention to HAR in cell xenotransplantation, especially differentiation ES cell in vivo.

筛选过程中在选择性培养基中添加LIF和BMP一2替代滋养层细胞，发现ES细胞的生长没有受到影响，提示在做基因敲除胚胎干细胞的筛选中可以不需要制备Nco抗性的滋养层细胞。

In the "Filter Action" tab, the "Use Add Wizard" removed from the left hook, click the "Add" button, add the "stop" operation: the "New Filter Action Properties" and "security" option card, select "stop", then click "OK" button.

在&筛选器操作&选项卡中，把&使用添加向导&左边的钩去掉，点击&添加&按钮，添加&阻止&操作：在&新筛选器操作属性&的&安全措施&选项卡中，选择&阻止&，然后点击&确定&按钮。

In the "Filter Action" tab, the "Use Add Wizard" removed from the left hook, click the "Add" button, add the "stop" operation: In the "New Filter Action Properties" of the "safe measures "tab, select the" stop ", then click" OK "button.

在&筛选器操作&选项卡中，把&使用添加向导&左边的钩去掉，点击&添加&按钮，添加&阻止&操作：在&新筛选器操作属性&的&安全措施&选项卡中，选择&阻止&，然后点击&确定&按钮。

The inconsistency of judgment matrixes, great difference of relative weights and other problems often arise during screening evaluation index by AHP. To solve these problems, a new AHP method is presented by combining the weight vector rang estimation analytic theory with traditional evaluation index rejecting method. Finally, an example of deciding a evaluation index system for submarine pipeline engineering project is given and the result proves that the method is effective and feasible.

针对用层次分析法进行评价指标筛选过程中经常出现比较矩阵不一致、相对权重值差别较大等问题，将权重向量区间估计分析理论与传统的评价指标剔除方法相结合，提出了一种新的AHP评价指标筛选方法，并以海底管道工程方案评价指标体系的确定为例进行了实际应用，结果表明该方法具有较强的实用性和可操作性。

Methods: With the Child Behavior Checklist and Conners Parent Symptom Questionnaire, two groups of students, including high aggression group and low aggression group, were selected from 311 primary school students, and the two groups were tested with the Rorschaeh test.

用Achenbach儿童行为量表和Conners教师问卷对湖南某地一小学311名同学进行筛选，筛选出攻击高分组(N=33)与低分组(N=35)，对这两组学生进行罗夏测验，回收有效数据，得到最后的高分组(N=30)和低分组(N=30)。

Therefore, we used a computer-based high throughput screening method to screen drug library to identify the possible binding drugs, thus increasing the added value for nsLTPs in drug delivery systems.

我们利用了电脑助药物筛选的技术来进行筛选，试找出可能与此一蛋白结合的药物，并结合萤光光谱的方法加以验证。如此，可进一步提高此一蛋白质的附加价值。

Keywords: NMT; Randomly phage display library; Affinity binding; Screening of inhibitor peptides; Molecule-screening model

中文关键词：豆蔻酰转移酶；噬菌体展示随机肽库；亲和结合；抑制肽筛选；分子筛选模型

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转印条件可调，能在很大的分子量跨度上获得最佳转印效果*耐用的板式电极能产生强大而均一的电场，而且电极在槽内的位置灵活可换；不论运行1、2 或3 个转印夹，电极间距可调节到最小以获得最大的场强和转印效率*凝胶支架转印夹保证整个凝胶与印迹膜表面的均衡接触*转印夹的铰链设计能防止凝胶滑动，便于转印夹组装*颜色标记的转印夹及电极板确保在转印槽内的正确定向*特级冷却芯和冷却水循环器进行温度调节―可理想地用于天然酶或高强度转印，或用于延长转印时间时减少缓冲液的损耗*可选择的组装盘是凝胶三明治和转印夹组装的理想选择 Trans-Blot Plus 转印槽能从单向和双向大格式凝胶上均一、高效地转移蛋白―配合使用新推出的 PowerPac HC 电源，多数蛋白都可在1530 分钟内完成转印。

On the other hand, the more important thing is because the urban housing is a kind of heterogeneity products.

另一方面，更重要的是由于城市住房是一种异质性产品。

Climate histogram is the fall that collects place measure calm value, cent serves as cross axle for a few equal interval, the area that the frequency that the value appears according to place is accumulated and becomes will be determined inside each interval, discharge the graph that rise with post, also be called histogram.

气候直方图是将所收集的降水量测定值，分为几个相等的区间作为横轴，并将各区间内所测定值依所出现的次数累积而成的面积，用柱子排起来的图形，也叫做柱状图。