Comparative genomic hybridization by array of barley BAC pools to anchor BACs physical and genetic maps
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ABSTRACT: BAC pool DNA hybridisation of barley to 44k Agilent microarrays. We have used two-channel Agilent expression microarrays to address thousands of gene sequences to individual BAC clones and contigs that form part of an emerging physical map of the large and unsequenced 5300 Mbp barley genome. By using two-colour processing, each array allows simultaneous co-hybridization of two independent BAC pools (SP), for which the data is analysed separately. As a general approach the method represents a cost-effective, highly parallel alternative to traditional gene-to-BAC addressing methods. By coupling the BAC address-data with gene-based genetic maps we were able to anchor thousands of BACs to the barley genetic map.
Project description:Background Because of its size, allohexaploid nature and high repeat content, the wheat genome has always been perceived as too complex for efficient molecular studies. However, we recently constructed the first physical map of a wheat chromosome (3B). But gene mapping is still laborious in wheat because of high redundancy between the three homoeologous genomes. In contrast, in the closely related diploid species, barley, numerous gene-based markers have been developed. This study aims at combining the unique genomic resources developed in wheat and barley to decipher the organisation of gene space on wheat chromosome 3B. Results Three dimensional pools of the minimal tiling path of wheat chromosome 3B physical map were hybridized to a barley Agilent 15K expression microarray. This led to the identification of 738 barley genes with a homolog on wheat chromosome 3B. In addition, comparative analyses revealed that 68% of the genes identified were syntenic between the wheat chromosome 3B and barley chromosome 3H and 59% between wheat chromosome 3B and rice chromosome 1, together with some wheat-specific rearrangements. Finally, it indicated an increasing gradient of gene density from the centromere to the telomeres positively correlated with the number of genes clustered in islands on wheat chromosome 3B. Conclusion Our study shows that novel structural genomics resources now available in wheat and barley can be combined efficiently to overcome specific problems of genetic anchoring of physical contigs in wheat and to perform high-resolution comparative analyses with rice for deciphering the organisation of the wheat gene space.
Project description:Profiling of DNA copy number alterations by array CGH in 100 sequential samples from 44 patients diagnosed with follicular lymphoma using a inhouse 1 Mb BAC/PAC array CGH
Project description:The huge size, the redundancy and the great repeated portion of the bread wheat genome [Triticum aestivum (L.)], placed it among the most difficult species to be sequenced and dissected at the genetic, structural and evolutionary levels. To overcome the limitations, a strategy based on the genome compartmentalization in individual chromosomes and the subsequent production of physical maps was established within the frame of the International Wheat Genome Sequence Consortium. A total of 95,812 BAC clones of short (5AS) and long (5AL) arm-specific BAC libraries, were fingerprinted and assembled into contigs by complementary analytical approaches based on FingerPrinted Contigs and Linear Topological Contig. Combined anchoring approaches based on PCR marker screening, microarray and BlastN searches, applied to interlinked genomic tools, that is genetic maps, deletion bin map, high-density neighbor map, BAC end sequences, genome zipper and chromosome survey sequences, allowed the development of a high quality physical map, with an anchored physical coverage of 75% for 5AS and 53% for 5AL, with high portions (64 and 48%, respectively) ordered along the chromosome. The gene distribution along the wheat chromosome 5A compared with the closest related genomes showed a pattern of syntenic blocks belonging to different chromosomes of Brachypodium, rice and sorghum and regions involving translocations and inversions. The physical map presented here is currently the most comprehensive map for 5A chromosome and represents an essential resource for fine genetic mapping and map-based cloning of agronomically relevant traits, and a reference for the 5A sequencing projects. 55 DNA pools of short arm of chromsome 5A and 63 DNA pools of long arm of 5A. The DNAs derive from BAC clones of the Minimal Tiling Paths produced by physical assemly of BAC fingerprints.
Project description:To obtain further insights into the role of bacterial activity in BAC filter performance, the expressed proteins of the bacterial community residing in the BAC filter were identified by a metaproteomic approach.
Project description:Ribonucleoprotein immunoprecipitation microarray (RIp-chip) study using various myosin proteins and other cytoskeletal components from the fission Schizosaccaromyces pombe
Project description:Lung cancer is the worldwide leading cause of death from cancer. This GEO series correspond to one of the BAC aCGH data sets used as validation cohort for the study: Landscape of somatic allelic imbalances and copy number alterations in human lung cancer, Int J Cancer 2012. Genomic profiling of 78 lung carcinomas using 32K BAC aCGH microarrays.
Project description:Transcriptomic profiling of laser microdissected epidermis and subepidermis cells from expanding green fruit rind of Citrus clemenules using the cDNA microarray CFGP-Citrus_20k.
Project description:To improve the resources for map-based cloning and sequencing of the wheat genome, we established a physical map of the wheat chromosome 1BL with a high density of markers by hybridizing the newly developed INRA GDEC Triticum aestivum NimbleGen 12x17k ISBP microarray (A-MEXP-2312) with BAC pools from the 1BL physical map. Then, we managed to map 3912 ISBP on the wheat chromosome 1BL BACs. The values in the 'Factor Value[individual]' column represent the BAC pool that have been hybridized on the array. For example, the assay 1 correspond to the hybridization of a bulk of all DNA BAC of the plate 1 of the MTP (Minimum Tilling path) BAC library of the chromosome 1BL.
Project description:Ribonucleoprotein immunoprecipitation microarray (RIp-chip) study using various myosin proteins and myosin-specific chaperones from the fission Schizosaccaromyces pombe