A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes

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 Hin...

Full description

Saved in:
Bibliographic Details
Published in:Theoretical and applied genetics 2000-11, Vol.101 (7), p.1093-1099
Main Authors: YU, Y, TOMKINS, J. P, WAUGH, R, FRISCH, D. A, KUDRNA, D, KLEINHOFS, A, BRUEGGEMAN, R. S, MUEHLBAUER, G. J, WISE, R. P, WING, R. A
Format: Article
Language:English
Subjects:
Artificial chromosomes
bacterial artificial chromosomes
Barley
Biological and medical sciences
Chloroplasts
Classical genetics, quantitative genetics, hybrids
Cloning
Corn
Crop diseases
Fundamental and applied biological sciences. Psychology
Genes
Genetics of eukaryotes. Biological and molecular evolution
Genomes
Genomics
Hordeum vulgare
Proteins
Pteridophyta, spermatophyta
Rice
Soil sciences
Sorghum
Vegetals
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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-cm^sup 2^ 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.[PUBLICATION ABSTRACT]
ISSN:0040-5752
1432-2242
DOI:10.1007/s001220051584