133 resultados para insertion


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报道了基于双面反射镜的N×N光开关器件。介绍了使用双面反射镜的2×2, 4×4光开关的集成光路设计和工作原理; 采用Benes网络, 以2×2和4×4光开关为基本单元的N×N光开关器件的整体结构, 并根据“一笔画”原理, 分析了4×4, 8×8和16×16光开关矩阵的可重排无阻塞特性和光开关矩阵的光路选择算法。最后, 基于2×2, 4×4光开关技术制备了16×16光开关矩阵。测试表明, 该器件具有良好的插入损耗、回波损耗、串扰和开关时间等性能, 从而验证了设计思想和工艺的可行性。在基于双面反射镜的光开关矩

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A wavelength division multiplexer (WDM) for 980/1550 nm based on planar curved waveguide coupler (CWC) is proposed. Compared with conventional parallel straight waveguide coupler (SWC), this structure has more flexibility with two variable parameters of bending radius R and minimum edge-to-edge spacing do, which are the two main parameters for the splitting ratio of coupler and decrease the complexity of device design and fabrication. Based on coupled mode theory (CMT) and waveguide theory, R and do of the WDM CWC are designed to be R = 13.28 m and d(0) = 4.39 mu m. The contrast ratio (CR) and insertion loss (IL) for 980 and 1550 nm are CR1 = 24.62 dB, CR2 = 24.56 dB and IL1 = 0.014 dB, IL2 = 0.015 dB, respectively. The 3D beam propagation method (BPM) is used to verify the validity of the design result. The influence of R and d(0) variations on the device performance is analyzed. For CR > 20 dB, the variation ranges of R and d(0) should be within -0.10 to +0.44 m and -0.05 to + 0.02 mu m, respectively. (c) 2006 Elsevier GmbH. All rights reserved.

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Effects of alumina and chromium interlayers on the microstructure and optical properties of thin Ag films are investigated by using spectrophotometry, x-ray diffraction and AFM. The characteristics of Ag films in Ag/glass, Ag/Al2O3/glass and Ag/Cr/glass stacks are analysed. The results indicate that the insertion of an Al2O3 or Cr layer decreases the grains and influences the reflectance of Ag films. The reflectance of the Ag film can be increased by controlling the thickness of alumina interlayer. The stability of Ag films is improved and the adhesion of Ag films on glass substrates is enhanced by alumina as an interlayer.

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拟南芥ast (anthocyanin spotted testa) 突变体是由碳离子束诱导产生的与花青苷生物合成有关的突变体,受单隐性核基因控制。由于花青苷的异常积累,突变体未成熟种子的种皮呈现紫红色的斑点;野生型植株幼嫩的种皮没有花青苷的异常积累,呈淡绿色。初步作图分析表明,AST基因定位于拟南芥第I号染色体上,并且位于SSLP分子标记nga280和CAPS分子标记PAB5之间。 AST基因与SSLP分子标记nga280紧密连锁,遗传距离为3.2cM;与CAPS分子标记PAB5相距较远,遗传距离为21.1cM。 采用DDRT-PCR的策略,分析野生型与突变型植株未成熟角果中基因表达的差异。通过调整DDRT-PCR中总RNA、锚定引物、随机引物、cDNA和dNTP等关键试剂的用量,优化了适用于银染检测的DDRT-PCR方法。PCR扩增产物经6%变性聚丙烯酰胺凝胶垂直电泳分离后,银染能检测到多而清晰的条带。泳道中的条带数最少为40个,最多达80个,平均为60个,条带大小分布在100bp-900bp范围,银染的灵敏度为5pg/mm2。此方法操作简便快速,灵敏度高,重复性好。采用这个改良的的方法,分析了拟南芥野生型和ast突变型植株未成熟角果中16,000个cDNA扩增产物条带,从中筛选出28个差异条带。二次PCR扩增后,进一步筛选出10个差异表达的cDNA条带,其中6个是野生型特异表达的,4个是突变型特异表达的。对这10个差异片段进行测序。BLASTN分析表明,这10个差异表达的cDNA片段与数据库中花青苷生物合成途径中的结构基因和调节基因序列没有同源性,表明用DDRT-PCR的方法克隆特定的AST基因有一定的局限性。 利用图位克隆(map-based cloning)的策略,对拟南芥AST 基因进行克隆。根据拟南芥数据库中的SNPs (simple nucleotide polymophisms) 序列和插入/缺失多态性(insertion/deletion polymorphisms)序列,设计了一系列分子标记。利用这些分子标记,对600个F2代有突变表型的植株进行重组子筛选,完成了对拟南芥AST基因的精细作图,成功地将AST 基因定位到BAC克隆T13M11上。初步确定该BAC克隆中的基因T13M11.8 可能是AST基因。该基因的DNA序列长1432bp,含有6个外显子和5个内含子,编码的蛋白与花青苷生物合成途径中的二氢黄酮醇4-还原酶有较高的同源性。功能互补实验正在进行当中。

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A bradykinin-like peptide has been isolated from the skin secretions of the frog Rana nigrovittata. This peptide was named ranakinin-N. Its primary structure, RAEAVPPGFTPFR, was determined by Edman degradation and mass spectrometry. It is structurally related to bradykinin-like peptides identified from skin secretions of other amphibians. Ranakinin-N is composed of 13 amino acid residues and is related to the bradykinin identified from the skin secretions of Odorrana schmackeri, which is composed of 9 amino acid residues. Ranakinin-N was found to exert concentration-dependent contractile effects on isolated guinea pig ileum. cDNA sequence encoding the precursor of ranakinin-N was isolated from a skin cDNA library of R. nigrovittata. The amino acid sequences deduced from the cDNA sequences match well with the results from Edman degradation. Analysis of different amphibian bradykinin cDNA structures revealed that the deficiency of a 15-nucleotide fragment (agaatgatcagacgc in the cDNA encoding bradykinin from O. schmackeri) in the peptide-coding region resulted in the absence of a dibasic site for trypsin-like proteinases and an unusual -AEVA- insertion in the N-terminal part of ranakinin-N. The -AEAV- insertion resulted in neutral net charge at the N-terminus of ranakinin-N. Ranakinin-N is the first reported bradykinin-like peptide with a neutral net charge at the N-terminus. Copyright (C) 2007 European Peptide Society and John Wiley & Sons, Ltd.

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A fragment of mitochondrial DNA (mtDNA) control region (similar to700 bp) was sequenced in 104 individuals from 20 breeds (three Chinese domestic breeds, five recently derived breeds and 12 introduced breeds) of domestic rabbits, Oryctolagus cuniculus . Nineteen sites were polymorphic, with 18 transitions and one insertion/deletion, and eight haplotypes (A1, A2, A3, A4, A5, A6, A7 and A8) were identified. Haplotype A1 was the most common and occurred in 89 individuals. In the 25 Chinese rabbits, only haplotype A1 was observed, while four haplotypes (A1, A3, A5 and A6) were found in 26 recently derived individuals. Haplotype A2 was shared by seven individuals among three introduced strains. The other six haplotypes accounted for 0. 96-1. 92% of the animals. Combined with the published sequences of European rabbits, a reduced median-joining network was constructed. The Chinese rabbit mtDNAs were scattered into two clusters of European rabbits. These results suggest that the (so-called) Chinese rabbits were introduced from Europe. Genetic diversity in Chinese rabbits was very low.

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An analysis of the nuclear beta-fibrinogen intron 7 locus from 30 taxa representing 12 placental orders of mammals reveals the enriched occurrences of short interspersed clement (SINE) insertion events. Mammalian-wide interspersed repeats (MIRs) are present at orthologous sites of all examined species except those in the order Rodentia. The higher substitution rate in mouse and a rare MIR deletion from rat account for the absence of MIR in the rodents. A minimum of five lineage-specific SINE sequences are also found to have independently inserted into this intron in Carnivora, Artiodactyla and Lagomorpha. In the case of Carnivora, the unique amplification pattern of order-specific CAN SINE provides important evidence for the "pan-carnivore" hypothesis of this repeat element and reveals that the CAN SINE family may still be active today. Particularly interesting is the finding that all identified lineage-specific SINE elements show a strong tendency to insert within or in very close proximity to the preexisting MIRs for their efficient integrations, suggesting that the MIR clement is a hot spot for successive insertions of other SINEs. The unexpected MIR excision as a result of a random deletion in the rat intron locus and the non-random site targeting detected by this study indicate that SINEs actually have a greater insertional flexibility and regional specificity than had previously been recognized. Implications for SINE sequence evolution upon and following integration, as well as the fascinating interactions between retroposons and the host genomes are discussed.

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Le polymorphisme au sein de quatre regions du gene codant pour la proteine prion bovine (PRNP) confere la susceptibilite a l'encephalopathie bovine spongiforme (BSE). Ceux-ci comprennent un polymorphisme d'insertion/deletion (indel) de 23 pb dans le promoteur, un indel de 12 pb dans l'intron 1, un octapeptide repete ou un indel de 24 pb au sein du cadre de lecture, et un polymorphisme mononucleotidique (SNP) dans la region codante. Dans ce travail, les auteurs ont examine la frequence des genotypes, des alleles et des haplotypes pour ces indel au sein de 349 bovins d'origine chinoise, de meme que la sequence nucleotidique de ce gene chez 50 de ces animaux. Leurs resultats montrent que l'allele ayant la deletion de 12 pb et l'haplotype combinant la deletion de 23 pb et la deletion de 12 pb, lesquels ont ete suggeres comme etant importants pour la susceptibilite a la BSE, sont rares au sein des bovins du sud de la Chine. Une difference significative a ete observee entre les bovins affectes par la BSE et les bovins chinois sains pour ce qui est de l'indel de 12 pb. Au total, 14 SNP ont ete observes dans la region codante du gene PRNP chez les bovins chinois. Trois de ces SNP etaient associes a des changements d'acides amines (K3T, P54S et S154N). La substitution E211K qui a ete rapportee recemment chez un cas atypique de la BSE chez un bovin americain n'a pas ete detectee dans ce travail.

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Objective: To investigate the association of complement C4 null genes (C4QO, including C4AQO and C4BQO) and C2 gene with systemic lupus erythematosus (SLE) in southwest Han Chinese; 136 patients with SLE and 174 matched controls were genotyped. Methods: C4 null genes were determined by a polymerase chain reaction (PCR) procedure with sequence specific primers (PCR-SSP). The 2 bp insertion in exon 29, which was previously identified in non-Chinese populations and caused defective C4A genes, was directly typed by sequencing the whole exon 29 using exon specific primers. The exon 6 of complement C2 was also sequenced in both the patients and controls. Results: The frequency of homozygous C4AQO allele was 12.5% (17/136) in patients with SLE compared with 1.1% (2/174) in controls (p<0.001, odds ratio (OR)=12.286, 95% confidence interval (95% CI) 2.786 to 54.170). There was no significant difference for homozygous C4BQO allele between patients with SLE and controls (p=0.699). Patients with the C4AQO gene had an increased risk of acquiring renal disorder, serositis, and anti-dsDNA antibodies compared with those without C4AQO (for renal disorder, p=0.018, OR=8.951, 95% Cl 1.132 to 70.804; for serositis, p=0.011, OR 4.891, 95% CI 1.574 to 15.198; for anti-dsDNA, p=0.004, OR 7.630, 95%Cl 1.636 to 35.584). None of the patients or controls had the 2 bp insertion in exon 29 of the C4 gene. The type I C2 deficiency was not detected in the 3 10 samples. Conclusion: It is suggested that deficiency of C4A (not due to a 2 bp insertion in exon 29), but not C4B or C2, may be a risk factor for acquiring SLE in south west Han Chinese; this results in increased risk of renal disorder, serositis, and anti-dsDNA antibodies in patients with SLE. Racial differences seem to be relevant in susceptibility to SLE.

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In plants and less-advanced animal species, such as C.elegans, introduction of exogenous double-stranded RNA (dsRNA) into cells would trigger degradation of the mRNA with homologous sequence and interfere with the endogenous gene expression. It might represent an ancient anti-virus response which could prevent the mutation in the genome that was caused by virus infection or mobile DNA elements insertion. This phenomenon was named RNA interference, or RNAi. In this study, RNAi was used to investigate the function of basonuclin gene during oogenesis. Microinjection of dsRNA directed towards basonuclin into mouse germinal-vesicle-intact (GV) oocytes brought down the abundance of the cognate mRNA effectively in a time- and concentration-dependent manner. This reduction effect was sequence-specific and showed no negative effect on other non-homologous gene expression in oocytes, which indicated that dsRNA can recognize and cause the degradation of the transcriptional products of endogenous basonuclin gene in a sequence-specific manner. Immunofluorescence results showed that RNAi could reduce the concentration of basonuclin protein to some extent, but the effect was less efficient than the dsRNA targeting towards tPA and cMos which was also expressed in oocytes. This result might be due to the long half life of basonuclin protein in oocytes and the short reaction time which was posed by the limited life span of GV oocytes cultured in vitro. In summary, dsRNA could inhibit the expression of the cognate gene in oocytes at both mRNA and protein levels. The effect was similar to Knock-out technique which was based on homologous recombination. Furthermore, hairpin-style dsRNA targeting basonuclin gene could be produced by transcription from a recombinant plasmid and worked efficiently to deplete the cognate mRNA in oocytes. This finding offered a new way to study the function of basonuclin in the early stage of oogenesis by infection of primordial oocytes with the plasmid expressing hairpin-style basonuclin dsRNA.

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Background: YidC/Oxa/Alb3 family includes a group of conserved translocases that are essential for protein insertion into inner membranes of bacteria and mitochondria, and thylakoid membranes of chloroplasts. Because mitochondria and chloroplasts are of b

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Polybrominated diphenyl ethers (PBDEs) are used extensively as flame-retardants and are ubiquitous in the environment and in wildlife and human tissue. Recent studies have shown that PBDEs induce neurotoxic effects in vivo and apoptosis in vitro. However, the signaling mechanisms responsible for these events are still unclear. In this study, we investigated the action of a commercial mixture of PBDEs (pentabrominated diphenyl ether, DE-71) on a human neuroblastoma cell line, SK-N-SH. A cell viability test showed a dose-dependent increase in lactate dehydrogenase leakage and 3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyl-tetrazolium bromide reduction. Cell apoptosis was observed through morphological examination, and DNA degradation in the cell cycle and cell apoptosis were demonstrated using flow cytometry and DNA laddering. The formation of reactive oxygen species was not observed, but DE-71 was found to significantly induce caspase-3, -8, and -9 activity, which suggests that apoptosis is not induced by oxidative stress but via a caspase-dependent pathway. We further investigated the intracellular calcium ([Ca2+](i)) levels using flow cytometry and observed an increase in the intracellular Ca2+ concentration with a time-dependent trend. We also found that the N-methyl d-aspartate (NMDA) receptor antagonist MK801 (3 mu M) significantly reduced DE-71-induced cell apoptosis. The results of a Western blotting test demonstrated that DE-71 treatment increases the level of Bax translocation to the mitochondria in a dose-dependent fashion and stimulates the release of cytochrome c (Cyt c) from the mitochondria into the cytoplasm. Overall, our results indicate that DE-71 induces the apoptosis of ([Ca2+](i)) in SK-N-SH cells via Bax insertion, Cyt c release in the mitochondria, and the caspase activation pathway.

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Pseudobagrus brachyrhabdion sp. nov., from the Yuan Jiang and Xiang Jiang of the middle Yangtze River drainage in Hunan and Guizhou Provinces, South China, is described herein. It is distinguished from all other Pseudobagrus species with a truncate or slightly emarginated caudal fin by an unique combination of the following characters: supraoccipital plate and nuchal plate broadly interspaced and covered with skin; nasal barbels only at most reaching anterior margin of eye; maxillary barbels reaching slightly beyond posterior margin of eye; outer mandibular barbels extending to posterior margin of eye; dorsal fin with a somewhat convex distal margin, origin nearer to pectoral-fin insertion than to pelvic-fin insertion; dorsal-fin spine shorter than pectoral spine, with a somewhat serrated posterior margin; pectoral-fin spine with a smooth anterior margin; anal fin with 20-23 rays, base length 23.8-32.0% of standard length, posterior end of anal-fin base anterior to posterior end of adipose fin base; no longitudinal black band extending along flank; eyes large, diameter 16.3-23.7% of head length; and number of vertebrae 5 + 43-46.

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Methyl parathion hydrolase (MPH) is an enzyme that catalyzes the degradation of methyl parathion, generating a yellow product with specific absorption at 405 nm. The application of MPH as a new labeling enzyme was illustrated in this study. The key advantages of using MPH as a labeling enzyme are as follows: (1) unlike alkaline phosphatase (AP), horseradish peroxidase (HRP), and glucose oxidase (GOD), MPH is rarely found in animal cells, and it therefore produces less background noise; (2) its active form in solution is the monomer, with a molecular weight of 37 kDa; (3) its turnover number is 114.70 +/- 13.19 s(-1), which is sufficiently high to yield a significant signal for sensitive detection; and (4) its 3D structure is known and its C-terminal that is exposed to the surface can be easily subjected to the construction of genetic engineering monocloning antibody-enzyme fusion for enzyme-linked immunosorbent assay (ELISA). To demonstrate its utility, MPH was ligated to an single-chain variable fragment (scFv), known as A1E, against a white spot syndrome virus (WSSV) with the insertion of a [-(Gly-Ser)(5)-] linker peptide. The resulting fusion protein MPH-A1E possessed both the binding specificity of the scFv segment and the catalytic activity of the MPH segment. When MPH-A1E was used as an ELISA reagent, 25 ng purified WSSV was detected; this was similar to the detection sensitivity obtained using A1E scFv and the HRP/Anti-E Tag Conjugate protocol. The fusion protein also recognized the WSSV in 1 mu L hemolymph from an infected shrimp and differentiated it from a healthy shrimp.

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From a random insertion mutant library of Synechocystis sp. PCC 6803, a mutant defective in photoautotrophic growth was obtained. The interrupted gene was identified to be slr2094 (rbpl), which encodes the fructose-1,6-biphosphatase (FBPase)/sedoheptulose-1,7-biphosphatase (SBPase) bifunctional enzyme (F-I). Two other independently constructed slr2094 mutants showed an identical phenotype. The FBPase activity was found to be virtually lacking in an slr2094 mutant, which was sensitive to light under mixotrophic growth conditions. These results indicate that slr2094 is the only active FBPase-encoding gene in this cyanobacterium. Inactivation of photosystem II by interrupting psbB in slr2094 mutant alleviated the sensitiveness to light. This report provides the direct genetic evidence for the essential role of F-I in the photosynthesis of Synechocystis sp. PCC 6803. (c) 2007 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.