Whole-genome fractionation rapidly purifies DNA from centromeric regions.
The condensed centromeric regions of higher eukaryotic chromosomes contain satellite sequences, transposons and retroelements, as well as transcribed genes that perform a variety of functions. These chromosomal domains nucleate kinetochores, mediate sister chromatid cohesion and inhibit recombination, yet their characterization has often lagged behind that of chromosome arms. Here, we describe a whole-genome fractionation technique that rapidly identifies bacterial artificial chromosome (BAC) clones derived from plant centromeric regions. This approach, which relies on hybridization of methylated genomic DNA, revealed BACs that correspond to the genetically mapped and sequenced Arabidopsis thaliana centromeric regions. Extending this method to other species in the Brassicaceae family identified centromere-linked clones and provided genome-wide estimates of methylated DNA abundance. Sequencing these clones will elucidate the changes that occur during plant centromere evolution. This genomic fractionation technique could identify centromeric DNA in genomes with similar methylation and repetitive DNA content, including those from crops and mammals.
Luo
Song
S
Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, Illinois 60637, USA.
Hall
Anne E
AE
Hall
Sarah E
SE
Preuss
Daphne
D
D017360:Arabidopsis / Q000235:genetics*; D002478:Cells, Cultured; D002503:Centromere / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D053938:DNA Fragmentation / Q000235:genetics*; D019175:DNA Methylation; D018744:DNA, Plant / Q000235:genetics* / Q000302:isolation & purification*; D018745:Genome, Plant; D017422:Sequence Analysis, DNA / Q000379:methods
D018744:DNA, Plant