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Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents

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Abstract

We have constructed a yeast artificial chromosome (YAC) library using high-molecular-weight DNA prepared from agarose-embedded leaf protoplasts of the maize inbred line UE95. This library contains 79 000 clones with an average insert size of 145 kb and should therefore represent approximately three haploid genome equivalents. The library is organised as an ordered array in duplicate microtitre plates. Forty-one pools of DNA from 1920 individual clones have been prepared for rapid screening of the library by the polymerase chain reaction (PCR). Using this approach, together with conventional colony hybridisation, we have been able to identify between one and eight positive clones for every probe used.

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References

  1. Albertsen HM, Abderrahim H, Cann HM, Dausset J, Paslier DL, Cohen D: Construction and characterisation of a yeast artificial chromosome library containing seven haploid human gene equivalents. Proc Natl Acad Sci USA 87: 4256–4260 (1990).

    PubMed  Google Scholar 

  2. Anand R, Villasante A, Tyler-Smith C: Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis. Nucl Acids Res 17: 3425–3432 (1989).

    PubMed  Google Scholar 

  3. Anand R, Riley JH, Butler R, Smith JC, Markham AF: A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage. Nucl Acids Res 18: 1951–1956 (1990).

    PubMed  Google Scholar 

  4. Bedbrook JR, Coen DM, Beaton AR, Bogorad L, Rich A: Location of the single gene for the large subunit of ribulose bisphosphate carboxylase on the maize chloroplast chromosome. J Biol Chem 254: 905–910 (1979).

    PubMed  Google Scholar 

  5. Bennett MD, Smith JB: Nuclear DNA amounts in Angiosperms. Trans Royal Society 274: 227–274 (1976).

    Google Scholar 

  6. Beverley SM: Characterisation of the unusual mobility of large circular DNA's in pulsed field-gradient electrophoresis. Nucleic Acids Res 16: 925–939 (1988).

    PubMed  Google Scholar 

  7. Brownstein BH, Silverman GA, Little RD, Burke T, Korsmeyer SJ, Schlessinger D, Olson MV: Isolation of single-copy human genes from a library of yeast artificial chromosome clones. Science 244: 1348–1351 (1989).

    PubMed  Google Scholar 

  8. Burke DT, Carle GF, Olson MV: Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236: 806–812 (1987).

    PubMed  Google Scholar 

  9. Collins FS: Chromosome jumping. In: Davies KE (ed.) Genome Analysis, pp. 78–79. IRL Press, Oxford.

  10. Freinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    PubMed  Google Scholar 

  11. Green ED, Olsen MV: Systematic screening of yeast artificial-chromosome libraries by use of the polymerase chain reaction. Proc Natl Acad Sci USA 87: 1213–1217 (1990).

    PubMed  Google Scholar 

  12. Grill E, Somerville C: Construction and characterisation of a yeast artificial chromosome library of Arabidopsis which is suitable for chromosome walking. Mol Gen Genet 226: 484–490 (1991).

    Article  PubMed  Google Scholar 

  13. Kao KN, Michayluk MR: Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very population density in liquid media. Planta 126: 105–110 (1975).

    Google Scholar 

  14. Knop W: Quantitative Untersuchungen über die Ernahrungsprozesse der Pflanzen. Landwirtsch Vers Stn 7: 93–107 (1865).

    Google Scholar 

  15. Martinez-Zapater JM, Estelle MA, Somerville CR: A highly repeated DNA sequence in Arabidopsis thaliana. Mol Gen Genet 204: 417–423 (1986).

    Article  Google Scholar 

  16. McIntosh L, Poulsen C, Bogorad L: Chloroplast gene sequence for the large subunit of ribulose bisphosphate-carboxylase of maize. Nature 288: 556–560 (1980).

    Google Scholar 

  17. Peacock WJ, Dennis ES, Rhoades MM, Pryor AJ: Highly repeated DNA sequence limited to knob heterochromatin in maize. Proc Natl Acad Sci USA 78: 4490–4494 (1981).

    Google Scholar 

  18. Sachs MM, Dennis ES, Gerlach WL, Peacock WJ: Two alleles of maize Alcohol dehydrogenase 1 have 3′ structural and poly(A) addition polymorphisms. Genetics 113: 449–467 (1986).

    Google Scholar 

  19. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).

    Google Scholar 

  20. Southern EM: Detection of specific sequences among DNA fragments separated gel electrophoresis. J Mol Biol 98: 503–517 (1975).

    PubMed  Google Scholar 

  21. Twigg AJ, Sherratt D: Trans-complementable copy number mutants of plasmid colE1. Nature 283: 216–218 (1980).

    PubMed  Google Scholar 

  22. Verdel F, Delseny M: Repetitivity and variability of higher plant genomes. Plant Physiol Biochem 25: 191–210 (1987).

    Google Scholar 

  23. Wahl GM, Stern M, Stark GR: Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl paper and rapid hybridisation using dextran sulfate. Proc Natl Acad Sci USA 76: 3683–3688 (1979).

    PubMed  Google Scholar 

  24. Ward ER, Jen GC: Isolation of single-copy-sequence clones from a yeast artificial chromosome library of randomly sheared Arabidopsis thaliana DNA. Plant Mol Biol 14: 561–568 (1990).

    PubMed  Google Scholar 

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Edwards, K.J., Thompson, H., Edwards, D. et al. Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents. Plant Mol Biol 19, 299–308 (1992). https://doi.org/10.1007/BF00027351

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  • DOI: https://doi.org/10.1007/BF00027351

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