Abstract
In the construction and characterization of yeast artificial chromosome (YAC) contigs, it is necessary to be able to isolate and map the ends of the genomic inserts. This is important with respect to both extending contigs, and identifying chimerism. A number of techniques have now been described and successfully applied to this purpose, and the majority of these methods are PCR-based, including Alu-vector PCR (1), vectorette libraries (2), and inverse PCR (3) (see ref. 4 for review). Although the possibility of isolating vector—insert junctions by plasmid rescue was identified in the initial article describing the pYAC vectors (5), these vectors were not designed with plasmid end rescue in mind. The most widely used YAC libraries are constructed in the pYAC4 vector, i.e., the Washington University (6), CEPH (7), ICRF (8), and ICI (9) libraries, and consequently, modifications to the vector arms need to be made in order to comprehensively use the plasmid rescue approach.
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References
Nelson, D. L., Ballabio, A., Victoria, M. F., Pieretti, M., Bies, R. D., Gibbs, R. A., et al. (1991) Alu-primed polymerase chain reaction for regional assignment of 110 yeast artificial chromosome clones from the human X chromosome: identification of clones associated with a disease locus. Proc. Natl. Acad. Sci. USA 88, 6157–6161.
Riley, J, Butler, R., Ogilvie, D., Finniear, R., Jenner, D., Powell, S., et al. (1990) A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones. Nucleic Acids Res. 18, 2887–2890.
Silverman, G. A., Ye, R. D., Pollock, K. M., Sadler, J. E., and Korsmeyer, S. J. (1989) Use of yeast artificial chromosome clones for mapping and walking within human chromosome segment 18q21 3. Proc. Natl.Acad. Sci. USA 86, 7485–7489.
Silverman, G A. (1993) Isolating vector—insert junctions from yeast artificial chromosomes. PCR Methods Appl. 3, 141–150.
Burke, D. T., Carle, G. F., and Olson, M. V. (1987) Cloning of large DNA segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236, 806–812.
Brownstein, B. H., Silverman, G. A., Little, R. D., Burke, D. T., Korsmeyer, S. J., Schlessinger, D., and Olson, M. V. (1989) Isolation of single-copy human genes from a library of yeast artificial chromosome clones. Science 244, 1348–1351.
Albertson, H. M., Abderrahim, H., Cann, H. M., Dausset, J., Paslier, D. L., and Cohen, D. (1990) Construction and characterisation of a yeast artificial chromosome library containing seven haploid human genome equivalents. Proc. Natl. Acad. Sci. USA 87, 4256–4260.
Larin, Z., Monaco, A. P., and Lehrach, H. (1991) Yeast artificial chromosome libraries containing large inserts from mouse and human DNA. Proc. Natl. Acad. Sci. USA 88, 4123–4127.
Anand, R., Riley, J. H., Butler, R., Smith, J. C., and Markham, A. F. (1990) A 3.5 genome equivalent multi access YAC library: construction, characterisation, screening and storage. Nucleic Acids Res. 18, 1951–1956.
Hermanson, G. G., Hoekstra, M. F., McElligot, D. L., and Evans, G. A. (1991) Rescue of end fragments of yeast artificial chromosomes by homologous recombination in yeast. Nucleic Acids Res. 19, 4943–4948.
Rose, M., Grisafi, P., and Botstein, D. (1984) Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene 29, 113–124.
Herrmann, B. G., Barlow, D. P., and Lehrach, H. (1987) An inverted duplication of more than 650 Kbp in mouse chromosome 17 mediates unequal but homologous recombination between chromosomes heterozygous for a large inversion. Cell 48, 813–825.
Church, G. M., and Gilbert, W. (1984) Genomic sequencing. Proc. Natl. Acad. Sci. USA 81, 1991–1995.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
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© 1996 Humana Press Inc.
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Bates, G. (1996). Isolation of YAC Ends by Plasmid Rescue. In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:139
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DOI: https://doi.org/10.1385/0-89603-313-9:139
Publisher Name: Humana Press
Print ISBN: 978-0-89603-313-9
Online ISBN: 978-1-59259-541-9
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