Abstract
For the isolation of gene space representative sequence elements, a new methodology—high C+G patch (HCGP) filtration—has been developed using rice as a model. The method is based on the fragmentation of the genomic DNA by methylation-sensitive HpaII and MspI restriction endonucleases having exclusively G/C base pair-containing recognition sites. These enzymes fragment the genome at high C+G content and hypomethylated regions. Cloning fragments spanning such regions in close vicinity (200–2,000 bp) revealed that about 60% of the clones represented gene space sequences resulting in twofold enrichment of these sequences, which is close to the theoretical maximum in rice.
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The sequence information of clones used in the present study was deposited in the NCBI database under the accession numbers EI 365676–EI 366364.
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Berenyi, M., Mauleon, R.P., Kopecky, D. et al. Isolation of Plant Gene Space-Related Sequence Elements by High C+G Patch (HCGP) Filtration: Model Study on Rice. Plant Mol Biol Rep 27, 79–85 (2009). https://doi.org/10.1007/s11105-008-0063-2
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DOI: https://doi.org/10.1007/s11105-008-0063-2