全长的

They conta i ned full genes ORFs and proved to have been prolongated effectively. The result showed that the seqences of 14 genes were genes known previously and 6 genes were new .We separately chose one seqence from known and the other from unknown genes to make full-length cDNA annotation. Their code number was EST contig110 and EST contig133. We designed specific primers to amplify the two genes by RT-PCR, and then sequenced. The result was obtained as expected and proved that silico cloning genes was reliable.

对20条contigs进行了基因注释，证实它们均得到了有效延伸，具有完整开放阅读框，其中已知的猪基因序列14条，猪新基因序列6条，分别从已知猪基因序列和未知猪基因序列中各抽取一条感兴趣序列进行全长cDNA的预测，它们的编号分别为EST contig110和EST contig133，设计特异引物对两条候选序列进行克隆与测序，结果与电子克隆所得序列一致，证实了电子克隆的有效性。

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 middle part of the sculpture is the map of the Qing Dynasty; another position at Dong'an Men (Dong'an Gate) is the historic site of Dong'an Men. Consisted of sunk squares, it has an area of 1,402 square meters; At the key point in the traditional courtyard house there stand a copper-coated sculpture named "Fashion" with 2.2 meters in height and 1.2 ton in weight and another called "Playing Chess", which is 2.2 meters high and 1 ton heavy

五四路口地下通道设浮雕11块，材质为玻璃钢，描述了东城区内的重要景点；中法大学节点面积300平方米；今天我们开会的茶室是另外一处重要景点，面积330平方米；公园北端点广场面积2100平方米，广场上恢复的皇城墙长25米，高6米，厚2.1米，公园从灯市西口至翠花胡同的地段，设立涌泉10个，全长400米，单个涌泉最长5米，高1.7米。

Methods cDNA retro-transcribed from equal quantities of mRNA derived from high and low metastatic tumor cells or normal ovarian tissues were labeled with Cy5 and Cy3 fluorescein as probes. The mixed probe was hybridized with two pieces of BioDoor 4096 double dot human whole gene chip and scanned with a ScanArray 3000 laser scanner. The acquired image was analyzed by ImaGene 3.0 software.

按一步法分别抽提高低转移人卵巢癌细胞和对照正常卵巢组织的总RNA并纯化mRNA ；分别将等量的高低转移人卵巢癌细胞及对照组织的mRNA逆转录合成以Cy5和Cy3标记的cDNA一链做探针，分别混合后在 2张含有 4 0 96条双点人类全长基因的芯片上进行杂交。

For the Cranberries, assembling a best-of covering the first decade of their career means compiling their singles in chronological order, which turns out to be a foolproof approach for a band that can be uneven over the course of a full-length album but has always managed to express itself well on the two-to-four catchiest cuts.

为蔓越桔，聚集的a 佳报道第一十年他们事业手段编写他们选拔在按年代顺序，哪些结果是一种极简单方法为可能是参差不齐的在全长册页中但的带总设法自我表达很好在二对四最吸引人的裁减。

The four proper complete DNA sequences were obtained by splicing the sequenced results with the software SeqMan of the DNA Star. The homology compare of the corresponding nuclear sequences and the cladogram analyze between the obtained virus strain and abroad virus collate strain were measured by Jotun Hein method in software DNA Star.

将MS-1、DL-1、DL-2和DL-3株病毒经组织病料提取总DNA，采用依据标准毒株ADV-G的DNA序列设计并合成的四对引物，经PCR扩增后分别构建重组质粒并测序，将测序结果用DNA Star软件中的SeqMan程序进行拼接，获得四株病毒的近全长DNA序列。

T3 is one of the worlds largest enclosed spaces, designed to handle an estimated 50 million visitors a year: It is just over two miles long, with 101 gates, 31 miles of baggage conveyers, 98 moving walkways, 168 escalators and 179 elevators.

福斯特设计的北京首都国际机场新航站楼那宏伟的穹拱形屋顶。3号航站楼是世界上最大的封闭空间之一，年接待量预计将达5，000万人次：航站楼全长约两英里，有101扇门，31英里长的行李传送带，98条电梯步道，168部自动扶梯和179部电梯。

T3 is one of the world's largest enclosed spaces, designed to handle an estimated 50 million visitors a year: It is just over two miles long, with 101 gates, 31 miles of baggage conveyers, 98 moving walkways, 168 escalators and 179 elevators.

福斯特设计的北京首都国际机场新航站楼那宏伟的穹拱形屋顶。3号航站楼是世界上最大的封闭空间之一，年接待量预计将达5，000万人次：航站楼全长约两英里，有101扇门，31英里长的行李传送带，98条电梯步道，168部自动扶梯和179部电梯。

To further elucidate the mechanism of BMP in regulating cell differentiation and to provide basis for BMP gene therapy, this study centers on the effects of BMP2 gene transfection on fibroblasts and contains the following works:① A 1.2kb fragment containing full length human BMP2 coding region was inserted into pBK-CMV (expressed both in protocaryon and eukaryon) by using predetermined orientation methods, resulting in phagemid expression vector for BMP2 (pBK-B2).② pBK-B2 was transfected into NIH3T3 cells by using LipofectAMINE.

为了进一步从分子水平探讨BMP对细胞分化的调控作用，并为BMP基因疗法的建立提供依据，本实验围绕BMP2基因转染对成纤维细胞的生物学作用进行了以下几个方面的工作：①将含有人BMP2全长编码序列的基因片段(1.2kb)定向克隆于噬菌粒表达载体pBK-CMV中，成功构建了人BMP2噬菌粒表达载体pBK-B2。

Longan1 expressed from the embryogenic callus stage to the globular embryoid stages. Longan2 expressed in the whole process of longan somatic embryogenesis, which was probably a housekeeping gene. Longan3 expressed from the globular embryoid stage to the torpedo-shaped embryoid stage. Longan4、longan4'expressed only at the cotyledonary embryoid stage.

碱基序列分析表明，longan1在3'端存在一个开放阅读框，longan2、longan3分别在5'端存在一个开放阅读框，longan1在3'方向，longan2、longan3在5'方向均没有终止密码子，所以可能往相应的方向延伸仍为各自的ORF，因此可以通过RACE或者筛选cDNA文库的办法来获得它们的全长cDNA。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。