Abstract
Recombinant DNA libraries were constructed for seven chromosome types isolated from two translocation lines of field bean (Vicia faba L.) with reconstructed karyotypes. The chromosomes were selected so that the set of libraries covers the wholeV. faba genome more than once. Individual chromosome types were highly purified by flow sorting, and their DNA was amplified by degenerate oligonucleotideprimed (DOP) polymerase chain reaction (PCR) and cloned into a plasmid vector. The choice of restriction site present in PCR primer and refinement of cloning protocol resulted in high cloning efficiency and allowed generation of libraries consisting of about 106 clones from 250 or 1000 sorted chromosomes. The insert size ranged between 50 and 2200 bp and the mean length estimated in individual libraries varied between 310 and 487 pb. Hybridization of cloned fragments with labelled genomic DNA showed that about 60% of inserts represented unique or low-copy sequences. The suitability of the libraries for genome mapping was demonstrated by isolation of clones containing microsatellite motifs.
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References
Arumuganathan K, Martin GB, Telenius H, Tanksley SD, Earle ED (1994) Chromosome 2-specific DNA clones from flow-sorted chromosomes of tomato.Mol Gen Genet 242: 551–558.
Ausbel FM, Brent R, Kingston RE et al. (1991)Current Protocols in Molecular Biology. New York: John Wiley & Sons.
Bennett MD, Leitch I (1995) Nuclear DNA amounts in angiosperms.Ann Bot 76: 113–176.
Breneman JW, Ramsey MJ, Lee DA, Eveleth GG, Minkler JL, Tucker JD (1993) The development of chromosome-specific composite DNA probes for the mouse and their application to chromosome painting.Chromosoma 102: 591–598.
Crowe JS, Cooper HJ, Smith MA, Sims MJ, Parker D, Gewert D (1991) Improved cloning efficiency of polymerase chain reaction (PCR) products after proteinase K digestion.Nucleic Acids Res 19: 184.
Doležel J, Lucretti S (1995) High-resolution flow karyotyping and chromosome sorting inVicia faba lines with standard and reconstructed karyotypes.Theor Appl Genet 90: 797–802.
Doležel J, Binarová P, Lucretti S (1989) Analysis of nuclear DNA content in plant cells by flow cytometry.Biol Plant 31: 113–120.
Doležel J, Sgorbati S, Lucretti S (1992a) Comparison of three DNA fluorochromes for flow cytometric estimation of nuclear DNA content in plants.Physiol Plant 85: 625–631.
Doležel J, Čihalíková J, Lucretti S (1992b) A high-yield procedure for isolation of metaphase chromosomes from root tips ofVicia faba L.Planta 188: 93–98.
Doležel J, Lucretti S, Schubert I (1994) Plant chromosome analysis and sorting by flow cytometry.Crit Rev Plant Sci 13: 275–309.
Doležel J, Lucretti S, Macas J. (1995) Flow cytometric analysis and sorting of plant chromosomes. In: Brandham PE, Bennett MD, eds.Kew Chromosome Conference IV. Kew: Royal Botanic Gardens, pp 185–200.
Fuchs J, Schubert I (1995) Localization of seed protein genes on metaphase chromosomes ofVicia faba via fluorescencein situ hybridization.Chrom Res 3: 94–100.
Gray JW, Cram LS. (1990) Flow karyotyping and chromosome sorting. In: Melamed MR, Lindmo T, Mendelsohn ML, eds.Flow Cytometry and Sorting. 2nd edn, New York: Wiley-Liss, pp 503–529.
Guan XY, Meltzer PS, Trent JM (1994) Rapid generation of whole chromosome painting probes (WCPs) by chromosome microdissection.Genomics 22: 101–107.
Hoebee B, Destoppelaar JM, Suijkerbuijk RF, Monard S (1994) Isolation of rat chromosome-specific paint probes by bivariate flow sorting followed by degenerate oligonucleotide primed PCR.Cytogenet Cell Genet 66: 277–282.
Jung V, Pestka SB, Pestka S (1993) Cloning of polymerase chain reaction-generated DNA containing terminal restriction endonuclease recognition sites. In: Wu R, ed.Methods in Enzymology. San Diego: Academic Press, pp 357–362.
Langford CF, Telenius H, Carter NP, Miller NGA, Tucker EM (1992) Chromosome painting using chromosome-specific probes from flow-sorted pig chromosomes.Cytogenet Cell Genet 61: 221–223.
Lucretti S, Doležel J, Schubert I, Fuchs J (1993) Flow karyotyping and sorting ofVicia faba chromosomes.Theor Appl Genet 85: 665–672.
Macas J, Doležel J, Lucretti S et al. (1993a) Localization of seed protein genes on flow-sorted field bean chromosomes.Chrom Res 1: 107–115.
Macas J, Weschke W, Baumlein H et al. (1993b) Localization of vicilin genes via polymerase chain reaction on microisolated field bean chromosomes.Plant J 3: 883–886.
McCormick MK, Buckler A, Bruno W et al. (1993) Construction and characterization of a YAC library with a low frequency of chimeric clones from flow-sorted human chromosome 9.Genomics 18: 553–558.
Pich U, Houben A, Fuchs J, Meister A, Schubert I (1994) Utility of DNA amplified by degenerate oligonucleotide-primed PCR (DOP-PCR) from the total genome and defined chromosomal regions of field bean.Mol Gen Genet 243: 173–177.
Riess O, Siedlaczck I, Kredtke S, Melmer G, Epplen JT, Deaven LL (1994) Characterization of a human chromosome-4 flow-sorted cosmid library.Cytogenet Cell Genet 65: 238–242.
Riquet J, Milan D, Woloszyn N et al. (1995) A linkage map with microsatellites isolated from swine flow-sorted chromosome 11.Mammalian Genome 6: 623–628.
Sanger F, Nicklen D, Coulson AR (1977) DNA sequencing with chain terminating inhibitors.Proc Natl Acad Sci USA 74: 5463–5467.
Schubert I, Rieger R, Michaelis A (1986) Structural and numerical manipulation of theVicia faba karyotype: results and perspectives.Biol Zentbl 105: 9–17.
Telenius H, Carter NP, Bebb CE, Nordenskjold M, Ponder BAJ, Tunnacliffe A (1992) Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer.Genomics 13: 718–725.
Torres AM, Weeden NF, Martin A (1993) Linkage among isozyme, RFLP and RAPD markers inVicia faba.Theor Appl Genet 85: 937–945.
Van Dilla MA, Deaven LL (1990) Construction of gene libraries for each human chromosome.Cytometry 11: 208–218.
Wang ML, Leitch AR, Schwarzacher T, Heslop-Harrison JS, Moore G (1992) Construction of a chromosome-enrichedHpaII library from flow-sorted wheat chromosomes.Nucleic Acids Res 20: 1897–1901.
Yakura K, Tanifuji S (1983) Molecular cloning and restriction analysis of EcoRI-fragments ofVicia faba rDNA.Plant Cell Physiol 24: 1327–1330.
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accepted for publication by J. S. (Pat) Heslop-Harrison
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Macas, J., Gualberti, G., Nouzová, M. et al. Construction of chromosome-specific DNA libraries covering the whole genome of field bean (Vicia faba L.). Chromosome Res 4, 531–539 (1996). https://doi.org/10.1007/BF02261781
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DOI: https://doi.org/10.1007/BF02261781