查询词典 structural gene

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

This is a very specific form of gene therapy that, if successful, will circumvent one the main objections to current gene therapy methods; namely, current methods insert the unmutated copy of a gene randomly into the genome, and if the insertion point happens to be near an oncogene, the gene therapy will cure one disease but cause another.

这种基因疗法是非常非常精确的。如果成功，现行的所有基因疗法将&报废&。现在的基因疗法是将正常的基因备份随机的加入到基因组中。如果切入点不对，疾病仍然可能发生。通过Rosetta研究出来的方法可以避免这一弊端。

Of the 32 articles, 4 were about the selection of destination genes, 3 about the selection of target cells, 7 about the selection of gene expression vector, 3 about the methods of gene transfer, 4 about gene therapy combined with tissue engineering in treating bone defect, 5 about polygenes in treating bone defects, and 2 about the problems and prospects in the gene therapy of bone defect.

符合纳入标准的32篇文献中，4篇涉及目的基因的选择，3篇涉及靶细胞的选择，7篇涉及基因表达载体的选择，3篇涉及基因转移方法，4篇涉及基因治疗与组织工程结合治疗骨缺损，5篇涉及多基因联合治疗骨缺损，2篇涉及骨缺损基因治疗存在的问题与展望。

Four pairs of primes were synthesized to amplify Salmonella genus special genes such as hut gene(495 bp), hilA gene(490 bp), invA gene(284 bp) and hns gene(152 bp) of Salmonella, respectively. The PCR reaction condition was optimized.

近年来，分子生物学方法如聚合酶链式反应在沙门氏菌的检测上已得到较好应用[1-6]，大大缩短了检测时间，但各报道中所用的DNA提取方法、引物、PCR反应条件各异。

It could be useful for the further study of NS1 gene function. Method The NS1 gene of H5N1 subtype AIV was amplified by RT_PCR and cloned to pGEM_T_easy vector. The NS1 gene expressing plasmids was constructed by inserting the target gene fragments into pGEX_4T_1 vectors. The expression proteins were indued by IPTG and analysed by Western blot.

方法采用RT_PCR方法对H5N1亚型AIVNS1基因进行扩增，将PCR产物克隆于pGEM_T_easy载体，将该基因插入pGEX_4T_1中构建NS1基因原核表达载体，转化BL21大肠杆菌后，在IPTG诱导下表达NS1蛋白，Westernblot鉴定表达NS1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。

This paper pointed out that the essential of geoherbs is continuers quantities variation at population level, geoherb's populations are different in gene frequency with the others'; geohersm are quantitative trait loci controlled by multi-gene or combination with multiple-gene and major gene at individual level.

指出道地性的遗传本质在居群水平通常是个量变的过程，它与种内其他非道地药材区别主要表现为居群内基因型频率的改变；在个体水平表现为微效多基因控制的数量遗传，或是微效多基因和主基因联合控制的数量性状。

The possibility of researching gene mutation with the technique of gene chip was discussed. A new imagination to randomly determine DNA sequence in different living beings with the technique of gene chip was advanced. In this way, whether the encoding sequence gene in DNA of eukaryon organism has some regularity was studied.

进而提出新设想——采用基因芯片技术对不同生物群体随机抽样进行DNA测序，从而探讨在真核生物DNA分子链上编码序列遗传基因的变化是否有规则而非随机的，非编码序列遗传基因是否为生物进化过程中曾经有表达能力的结构基因，并分析可能获得的结果。

Bioinformatics was used to seek the homologous gene of EXT1 in Strongylocentrotus purpuratus which was just discovered and had the correlation to human hereditary multiple exostoses. The sequence, exons, coding protein, physical and chemical characters of EXT1 gene were analyzed. The structure and function of its coding protein were predicted, and the phylogenetic tree for the homologous gene was constructed, which provided certain basis for the future research of human EXT1 gene.

利用生物信息学方法寻找人类遗传多发性外生性骨疣EXT1基因在紫色球海胆中的同源基因，对该基因的序列、外显子信息、编码蛋白及其理化性质进行分析，并预测其编码蛋白的结构与功能，构建其同源基因的系统进化树，旨在为进一步研究人体EXT1基因提供一定的依据。

This expression distribution suggests that COCH gene may contribute to maintain inner ear connective tissue extracellular matrix architecture, and do not support the postulation that gene might play an inner ear immune defensive function. 2. In vitro, the culturing spiral ligament type Ⅰ fibrocytes expressed COCH gene and POU3F4/brn4 gene, which were checked by RT-PCR and Western blot, and expression level did not chang with in 15 generations.

小鼠内耳COCH基因原位表达分布的观察结果提示，基因的功能与维持内耳间质组织细胞外基质的结构稳定有关，不支持COCH基因的功能与内耳免疫应答反应有关的推论。

These are places without aristocratic baggage; egalitarian places open to talent, self-improving, engaged in learning and innovation through networks that were at once competitive and cooperative.

这些地方没有贵族遗风作祟，而且对于那些有天分的人是开放的平等之地，这些人善于通过那些曾经很具有竞争力与合作精神的关系网进行自我提高以及学习创新。

Christine: You don't want to see me?

你不想见我？