共查询到20条相似文献,搜索用时 15 毫秒
1.
De Preter K Pattyn F Berx G Strumane K Menten B Van Roy F De Paepe A Speleman F Vandesompele J 《BMC genomics》2004,5(1):11-14
Background
Activation of proto-oncogenes by DNA amplification is an important mechanism in the development and maintenance of cancer cells. Until recently, identification of the targeted genes relied on labour intensive and time consuming positional cloning methods. In this study, we outline a straightforward and efficient strategy for fast and comprehensive cloning of amplified and overexpressed genes. 相似文献2.
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Yongxin Wang Joseph M Fugaro Fauzia Siddiq Chandra Mouli V Goparaju Fulvio Lonardo Anil Wali John F Lechner Harvey I Pass 《BMC molecular biology》2000,1(1):2-4
Background
PCR amplification of target molecules involves sequence specific primers that flank the region to be amplified. While this technique is generally routine, its applicability may not be sufficient to generate a desired target molecule from two separate regions involving intron /exon boundaries. For these situations, the generation of full-length complementary DNAs from two partial genomic clones becomes necessary for the family of low abundance genes. 相似文献4.
Limei Hu Jing Wang Keith Baggerly Hua Wang Gregory N Fuller Stanley R Hamilton Kevin R Coombes Wei Zhang 《BMC genomics》2002,3(1):16-8
Background
High density cDNA microarray technology provides a powerful tool to survey the activity of thousands of genes in normal and diseased cells, which helps us both to understand the molecular basis of the disease and to identify potential targets for therapeutic intervention. The promise of this technology has been hampered by the large amount of biological material required for the experiments (more than 50 μg of total RNA per array). We have modified an amplification procedure that requires only 1 μg of total RNA. Analyses of the results showed that most genes that were detected as expressed or differentially expressed using the regular protocol were also detected using the amplification protocol. In addition, many genes that were undetected or weakly detected using the regular protocol were clearly detected using the amplification protocol. We have carried out a series of confirmation studies by northern blotting, western blotting, and immunohistochemistry assays. 相似文献5.
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Elena?Busti Roberta?Bordoni Bianca?Castiglioni Paolo?Monciardini Margherita?Sosio Stefano?Donadio Clarissa?Consolandi Luigi?Rossi Bernardi Cristina?Battaglia Gianluca?De Bellis
Background
PCR amplification of bacterial 16S rRNA genes provides the most comprehensive and flexible means of sampling bacterial communities. Sequence analysis of these cloned fragments can provide a qualitative and quantitative insight of the microbial population under scrutiny although this approach is not suited to large-scale screenings. Other methods, such as denaturing gradient gel electrophoresis, heteroduplex or terminal restriction fragment analysis are rapid and therefore amenable to field-scale experiments. A very recent addition to these analytical tools is represented by microarray technology. 相似文献8.
Alejandro Hernández-Morales Susana De la Torre-Zavala Enrique Ibarra-Laclette José Luis Hernández-Flores Alba Estela Jofre-Garfias Agustino Martínez-Antonio Ariel álvarez-Morales 《BMC microbiology》2009,9(1):257
Background
Pseudomonas syringae pv. phaseolicola is a Gram-negative plant-pathogenic bacterium that causes "halo blight" disease of beans (Phaseolus vulgaris L.). This disease affects both foliage and pods, and is a major problem in temperate areas of the world. Although several bacterial genes have been determined as participants in pathogenesis, the overall process still remains poorly understood, mainly because the identity and function of many of the genes are largely unknown. In this work, a genomic library of P. syringae pv. phaseolicola NPS3121 was constructed and PCR amplification of individual fragments was carried out in order to print a DNA microarray. This microarray was used to identify genes that are differentially expressed when bean leaf extracts, pod extracts or apoplastic fluid were added to the growth medium. 相似文献9.
Kung Ahn Jae-Won Huh Sang-Je Park Dae-Soo Kim Hong-Seok Ha Yun-Ji Kim Ja-Rang Lee Kyu-Tae Chang Heui-Soo Kim 《BMC molecular biology》2008,9(1):78
Background
The rhesus monkey (Macaca mulatta) is a valuable and widely used model animal for biomedical research. However, quantitative analyses of rhesus gene expression profiles under diverse experimental conditions are limited by a shortage of suitable internal controls for the normalization of mRNA levels. In this study, we used a systematic approach for the selection of potential reference genes in the rhesus monkey and compared their suitability to that of the corresponding genes in humans. 相似文献10.
M. Storari R. von Rohr I. Pertot C. Gessler G.A.L. Broggini 《Journal of applied microbiology》2013,114(4):1193-1200
Aims
To develop two assays based on the loop‐mediated isothermal amplification (LAMP) of DNA for the quick and specific identification of Aspergillus carbonarius and ochratoxigenic strains of the Aspergillus niger clade isolated from grapes.Methods and Results
Two sets of primers were designed based on the polyketide synthase genes involved or putatively involved in ochratoxin A (OTA) biosynthesis in A. carbonarius and A. niger clade. Hydroxynaphthol blue was used as indirect method to indicate DNA amplification. The limit of detection of both assays was comparable to that of a PCR reaction. Specificities of the reactions were tested using DNA from different black aspergilli isolated from grapes. The two LAMP assays were then used to identify A. carbonarius and ochratoxigenic A. niger and A. awamori grown in pure cultures without a prior DNA extraction.Conclusions
The two LAMP assays permitted to quickly and specifically identify DNA from OTA‐producing black aspergilli, as well as isolates grown in pure culture.Significance and Impact of the Study
Monitoring vineyards for the presence of OTA‐producing strains is part of the measures to minimize the occurrence of OTA in grape products. The two LAMP assays developed here could be potentially used to speed the screening process of vineyards for the presence of OTA‐producing black aspergilli. 相似文献11.
Morgane Thomas-Chollier Valérie Ledent Luc Leyns Michel Vervoort 《BMC evolutionary biology》2010,10(1):73
Background
Hox and the closely-related ParaHox genes, which emerged prior to the divergence between cnidarians and bilaterians, are the most well-known members of the ancient genetic toolkit that controls embryonic development across all metazoans. Fundamental questions relative to their origin and evolutionary relationships remain however unresolved. We investigate here the evolution of metazoan Hox and ParaHox genes using the HoxPred program that allows the identification of Hox genes without the need of phylogenetic tree reconstructions. 相似文献12.
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Kira S Makarova Alexander V Sorokin Pavel S Novichkov Yuri I Wolf Eugene V Koonin 《Biology direct》2007,2(1):33-20
Background
An evolutionary classification of genes from sequenced genomes that distinguishes between orthologs and paralogs is indispensable for genome annotation and evolutionary reconstruction. Shortly after multiple genome sequences of bacteria, archaea, and unicellular eukaryotes became available, an attempt on such a classification was implemented in Clusters of Orthologous Groups of proteins (COGs). Rapid accumulation of genome sequences creates opportunities for refining COGs but also represents a challenge because of error amplification. One of the practical strategies involves construction of refined COGs for phylogenetically compact subsets of genomes. 相似文献15.
Gerald Hochwimmer Reinhard Tober Renè Bibars-Reiter Elisabeth Licek Ralf Steinborn 《BMC microbiology》2009,9(1):184-17
Background
The oomycete Aphanomyces astaci is regarded as the causative agent of crayfish plague and represents an evident hazard for European crayfish species. Native crayfish populations infected with this pathogen suffer up to 100% mortality. The existence of multiple transmission paths necessitates the development of a reliable, robust and efficient test to detect the pathogen. Currently, A. astaci is diagnosed by a PCR-based assay that suffers from cross-reactivity to other species. We developed an alternative closed-tube assay for A. astaci, which achieves robustness through simultaneous amplification of multiple functionally constrained genes. 相似文献16.
Andreas V Hadjinicolaou Victoria L Demetriou Maria A Emmanuel Charalambos K Kakoyiannis Leondios G Kostrikis 《BMC microbiology》2009,9(1):97
Background
A fast and simple two-step multiplex real-time PCR assay has been developed to replace the traditional, laborious Salmonella serotyping procedure. Molecular beacons were incorporated into the assay as probes for target DNA. Target sequences were regions of the invA, prot6E and fliC genes specific for Salmonella spp. Salmonella Enteritidis and Salmonella Typhimurium, respectively, the two most clinically relevant serotypes. An internal amplification positive control was included in the experiment to ensure the optimal functioning of the PCR and detect possible PCR inhibition. Three sets of primers were used for the amplification of the target sequences. The results were compared to those of the Kauffmann-White antigenic classification scheme. 相似文献17.
Martijn F Schenk Jan HG Cordewener Antoine HP America Wendy PC van't Westende Marinus JM Smulders Luud JWJ Gilissen 《BMC plant biology》2009,9(1):24
Background
Bet v 1 is an important cause of hay fever in northern Europe. Bet v 1 isoforms from the European white birch (Betula pendula) have been investigated extensively, but the allergenic potency of other birch species is unknown. The presence of Bet v 1 and closely related PR-10 genes in the genome was established by amplification and sequencing of alleles from eight birch species that represent the four subgenera within the genus Betula. Q-TOF LC-MSE was applied to identify which PR-10/Bet v 1 genes are actually expressed in pollen and to determine the relative abundances of individual isoforms in the pollen proteome. 相似文献18.
Elisa Porcellini Ilaria Carbone Manuela Ianni Federico Licastro 《Immunity & ageing : I & A》2010,7(1):16
Background
Recent findings from a genome wide association investigation in a large cohort of patients with Alzheimer's disease (AD) and non demented controls (CTR) showed that a limited set of genes was in a strong association (p > l0-5) with the disease. 相似文献19.
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Huber R Hummert C Gausmann U Pohlers D Koczan D Guthke R Kinne RW 《Arthritis research & therapy》2008,10(4):R98