naa a22 4500 510730825 CHVBK 20180411082952.0 cr unu---uuuuu 180411e20130801xx s 000 0 eng 10.1007/s11295-013-0621-0 doi (NATIONALLICENCE)springer-10.1007/s11295-013-0621-0 Identification of Citrus sinensis BAC clones containing genes relevant to fruit quality by two-dimensional overgo hybridization [Elektronische Daten] [Sergio Ferrante, Mikeal Roose] As a complement to whole-genome sequencing efforts, we are comparing targeted genomic regions among sweet orange cultivars to identify coding and conserved noncoding regions, including regulatory elements, responsible for biological features unique to this species. Here, we report the identification of 1,018 bacterial artificial chromosome (BAC) clones containing genes relevant to fruit quality from a Citrus sinensis cv. "Vaniglia” 19.3X BAC library by two-dimensional 9 × 9 overgo hybridization. To design the overgo probes, we used the "C38” expressed sequence tag assembly ( http://harvest.ucr.edu/ ) and OligoSpawn software ( http://138.23.178.42 ). For BAC library screening, we selected 81 overgo probes associated with unigenes that putatively code for enzymes relevant to fruit quality (flavonol, anthocyanin, carotenoid, cellulose, starch, ascorbic acid, aromatic amino acid, and lignin biosynthesis; sucrose catabolism; glycolysis; oxidative/nonoxidative pentose phosphate pathway; fatty acid biosynthesis and oxidation; Krebs cycle). Hybridization probes were pooled and hybridized in groups of intersecting rows and columns to high-density BAC filters, followed by a deconvolution process that established BAC-probe addresses. BAC addresses were obtained for 75 of the 81 overgo probes initially selected, for a total of 1,018 BAC clones, a number consistent with the depth of coverage of the BAC library. BAC end sequencing was carried out, and end-sequence pairs were mapped to their best location in the Citrus clementina genome sequence assembly using the comparative genomic database Phytozome ( http://www.phytozome.net/ ). The BAC clones corresponding to each probe were mapped within the same scaffold as the target gene, demonstrating that the approach we used was successful in isolating the targeted genomic regions. Springer-Verlag Berlin Heidelberg, 2013 Unigene nationallicence Repeat masking nationallicence Pooling strategy nationallicence Deconvolution nationallicence Structural variation nationallicence Ferrante Sergio Institute of Plant Genetics, Section of Palermo, CNR—National Research Council of Italy, Corso Calatafimi 414, 90129, Palermo, Italy aut Roose Mikeal Department of Botany and Plant Sciences, University of California Riverside, 92521, Riverside, CA, USA aut Tree Genetics & Genomes Springer Berlin Heidelberg 9/4(2013-08-01), 1065-1074 1614-2942 9:4<1065 2013 9 11295 https://doi.org/10.1007/s11295-013-0621-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11295-013-0621-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ferrante Sergio Institute of Plant Genetics, Section of Palermo, CNR—National Research Council of Italy, Corso Calatafimi 414, 90129, Palermo, Italy aut NATIONALLICENCE 700 1- Roose Mikeal Department of Botany and Plant Sciences, University of California Riverside, 92521, Riverside, CA, USA aut NATIONALLICENCE 773 0- Tree Genetics & Genomes Springer Berlin Heidelberg 9/4(2013-08-01), 1065-1074 1614-2942 9:4<1065 2013 9 11295 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer