Abstract
A yeast artificial chromosome (YAC) genomic library of Beta vulgaris was constructed in the pYAC4 vector. High-molecular-weight DNA was prepared from agarose-embedded leaf protoplasts from a triploid cultivar. The library was found to contain 33,500 clones in an ordered array of microtiter plates. Mean size of the inserts was estimated to be 135 kb, and the library should therefore represent the equivalent of five haploid genomes. The library was characterised for the presence of highly repetitive, chloroplast and single-copy sequences. In order to isolate single-copy sequences, 18 pools of DNA, each from 1920 individual YAC clones, were prepared for rapid screening of the library by the polymerase chain reaction. The results of these screenings showed that the number of isolated clones was at or near the frequency expected.
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Albertsen HM, Abderrahim H, Cann HM, Dausset J, Le Paslier D, Cohen D (1990) Construction and characterization of a yeast artificial chromosome library containing seven haploid genome equivalents. Proc Natl Acad Sci USA 87:4256–4260
Anand R, Villasante A, Tyler-Smith C (1989) Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis. Nucleic Acids Res 17:3425–3432
Barzen E, Mechelke W, Ritter E, Seitzer JF, Salamini F (1992) RFLP markers for sugar beet breeding: chromosomal linkage maps and location of major genes for rhizomania resistance, monogermy and hypocotyl colour. Plant J 2:601–611
Brownstein BH, Silverman GA, Little RD, Burke T, Korsmeyer SJ, Schlessinger D, Olson MV (1989) Isolation of single-copy human genes from a library of yeast artificial chromosome clones. Science 244:1348–1351
Burgers PMJ, Percival KJ (1987) Transformation of yeast spheroplasts without cell fusion. Anal Biochem 163:391–397
Burke DT, Carle GF, Olsen MV (1987) Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236:806–812
Connelly C, McCormick MK, Shero J, Hieter P (1991) Polyamines eliminate an extreme size bias against transformation of large yeast artificial chromosome DNA. Genomics 10:10–16
Edwards KJ, Thompson H, Edwards D, de Saizieu A, Sparks C, Thompson JA, Greenland AJ, Eyers M, Schuch W (1992) Construction and characterization of a yeast artificial chromosome library containing three haploid genome equivalents. Plant Mol Biol 19:299–308
Eyers M, Edwards K, Schuch W (1992) Construction and characterization of a yeast artificial chromosome library containing two haploid Beta vulgaris L. equivalents. Gene 121:195–202
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Green ED, Olsen MV (1990) Systematic screening of yeast artificial-chromosome libraries by use of the polymerase chain reaction. Proc Natl Acad Sci USA 87:1213–1217
Grill E, Somerville C (1991) Construction and characterization of a yeast artificial chromosome library of Arabidiopsis which is suitable for chromosome walking. Mol Gen Genet 226:484–490
Larin Z, Monaco AP, Lehrach H (1991) Yeast artificial chromosome libraries containing large inserts from mouse and human DNA. Proc Natl Acad Sci USA 88:4123–4127
Martin GB, Ganal WM, Tanksley SD (1992) Construction of a yeast artificial chromosome library of tomato and identification of cloned segments linked to two disease resistance loci. Mol Gen Genet 233:25–32
McCue KF, Hanson AD (1992) Salt-inducible betaine aldehyde dehydrogenase from sugar beet: cDNA cloning and expression. Plant Mol Biol 18:1–11
McIntosh L, Poulsen C, Bogorad L (1980) Chloroplast gene sequence for the large subunit of ribulose biphosphate carboxylase of maize. Nature 288:556–560
Pillen K, Steinbrücken G, Wricke G, Herrmann RG, Jung C (1992) A linkage map of sugar beet Theor Appl Genet 84:129–135
Schlangstedt M, Hermans B, Zoglauer K, Schieder O (1992) Culture of sugar beet (Beta vulgaris L.) protoplasts in alginate — callus formation and root organogenesis. J Plant Physiol 140:339–344
Schmidt T, Metzlaff (1991) Cloning and characterization of a Beta vulgaris satellite DNA family. Gene 101:247–250
van Daelen RAJ, Jonkers JJ, Zabel P (1989) Preparation of megabase-sized tomato DNA and separation of large restriction fragments by field inversion gel electrophoresis (FIGE). Plant Mol Biol 12:341–352
Vedel F, Delseny M (1987) Repetitivity and variability of higher plant genomes. Plant Physiol Biochem 25:191–210
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Communicated by J. S. Beckmann
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Del-Favero, J., Vauterin, M., Weyens, G. et al. Construction and characterisation of a yeast artificial chromosome library containing five haploid sugarbeet (Beta vulgaris L.) genome equivalents. Theoret. Appl. Genetics 88, 449–453 (1994). https://doi.org/10.1007/BF00223659
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DOI: https://doi.org/10.1007/BF00223659