caa a22 4500 475793609 CHVBK 20180406123717.0 cr unu---uuuuu 170329e20001101xx s 000 0 eng 10.1007/s001220051584 doi (NATIONALLICENCE)springer-10.1007/s001220051584 A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes [Elektronische Daten] [Y. Yu, J. P. Tomkins, R. Waugh, D. A. Frisch, D. Kudrna, A. Kleinhofs, R. S. Brueggeman, G. J. Muehlbauer, R. P. Wise, R. A. Wing] Abstract : Modern cultivated barley is an important cereal crop with an estimated genome size of 5000 Mb. To develop the resources for positional cloning and structural genomic analyses in barley, we constructed a bacterial artificial chromosome (BAC) library for the cultivar Morex using the cloning enzyme HindIII. The library contains 313344 clones (816 384-well plates). A random sampling of 504 clones indicated an average insert size of 106 kbp (range=30-195 kbp) and 3.4% empty vectors. Screening the colony filters for chloroplast DNA content indicated an exceptionally low 1.5% contamination with chloroplast DNA. Thus, the library provides 6.3 haploid genome equivalents allowing a >99% probability of recovering any specific sequence of interest. High-density filters were gridded robotically using a Genetix Q-BOT in a 4×4 double-spotted array on 22.5-cm2 filters. Each set of 17 filters allows the entire library to be screened with 18432 clones represented per filter. Screening the library with 40 single copy probes identified an average 6.4 clones per probe, with a range of 1-13 clones per probe. A set of resistance-gene analog (RGA) sequences identified 121 RGA-containing BAC clones representing 20 different regions of the genome with an average of 6.1 clones per locus. Additional screening of the library with a P-loop disease resistance primer probe identified 459 positive BAC clones. These data indicate that this library is a valuable resource for structural genomic applications in barley. Springer-Verlag Berlin Heidelberg, 2000 Key words Barley nationallicence BAC library nationallicence P-loop genes nationallicence Resistance-gene analog (RGA) nationallicence Yu Y. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut Tomkins J. P. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut Waugh R. Scottish Crop Research Institute, Invergowrie, Dundee, DD2 2DA, UK, GB aut Frisch D. A. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut Kudrna D. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut Kleinhofs A. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut Brueggeman R. S. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut Muehlbauer G. J. Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA, US aut Wise R. P. USDA-ARS, Corn Insects and Crop Genetics Research Unit, Department of Plant Pathology, Iowa Stae University, Ames, IA 50011-1020, USA, US aut Wing R. A. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut https://doi.org/10.1007/s001220051584 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001220051584 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yu Y. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut NATIONALLICENCE 700 1- Tomkins J. P. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut NATIONALLICENCE 700 1- Waugh R. Scottish Crop Research Institute, Invergowrie, Dundee, DD2 2DA, UK, GB aut NATIONALLICENCE 700 1- Frisch D. A. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut NATIONALLICENCE 700 1- Kudrna D. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut NATIONALLICENCE 700 1- Kleinhofs A. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut NATIONALLICENCE 700 1- Brueggeman R. S. Department of Crop and Soil Sciences, Washington State University, Pullman, WA 99164, USA, US aut NATIONALLICENCE 700 1- Muehlbauer G. J. Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA, US aut NATIONALLICENCE 700 1- Wise R. P. USDA-ARS, Corn Insects and Crop Genetics Research Unit, Department of Plant Pathology, Iowa Stae University, Ames, IA 50011-1020, USA, US aut NATIONALLICENCE 700 1- Wing R. A. Clemson University Genomics Institute, BAC/EST Resource Center, Room 100 Jordan Hall, Clemson, SC 29634, USA e-mail: jtmkns@clemson.edu e-mail: rwing@clemson.edu, JO aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer