Abstract
We report the construction of a YAC library that provides 10-fold redundant coverage of the chicken genome. The library was made by transforming S. cerevisiae AB1380 with YAC constructs consisting of partially digested and size fractionated (>465 kb) EcoRI genomic fragments ligated to pCGS966 YAC vector arms. The primary library provides 8.5-fold redundant coverage and consists of 16,000 clones arrayed in duplicate 96-well microtiter plates and gridded on nylon membranes at high density (18,000 clones/484cm2). The average insert size, 634 kb, was derived from size fractionation of a random sample of 218 YACs. Hybridization of five unlinked chicken genes to colony blots revealed six or more positive clones. This is consistent with the theoretical expectation from average insert sizes and number of clones. A second collection of clones consists of a further 20,000 colonies, of which 20% contain inserts larger than 450 kb and 80% contain only coligated vector arms. We estimate that these clones provide a further 1.5-fold redundant coverage of the chicken genome; thus, the total collection of 36,000 clones provides 10-fold redundant coverage of the chicken genome. The library is intended as a resource for fine-scale analysis of the organization of the chicken genome and is presently being used to construct a contig map of chicken Chromosome (Chr) 16, which contains the MHC and nucleolar organizer.
Similar content being viewed by others
References
Albertsen HM, Abderrahim H, Cann HM, Dausset J, Le-Paslier D, 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
Ballabio A (1993) The rise and fall of positional cloning? Nature Genet 3, 277–279
Bumstead N (1996) Breeding for disease resistance. In Poultry Immunology ed. TF Davison, TR Mories, LN Payne. Carfax Publishing: Abingdon, UK pp 405–415
Bumstead N, Palyga J (1992) A preliminary linkage map of the chicken genome. Genomics 13, 690–697
Collins FS (1992) Positional cloning: let’s not call it reverse anymore. Nature Genet 1, 3–6
Den-Dunnen JT, Grootscholten PM, Dauwerse JG, Walker AP, Monaco AP, Butler R, Anand R, Coffey AJ, Bentley DR, Steensma HY, Van Ommen GJB (1992) Reconstruction of the 2.4 Mb human DMD-gene by homologous YAC recombination. Hum Mol Genet 1, 19–28
Deitrich WF, Copeland NF, Gilbert DJ, Miller JC, Jenkins NA, Lander ES (1995) Mapping the mouse genome: current status and future prospects. Proc Natl Acad Sci USA 92, 10849–10853
Heale SM, Stateva LI, Oliver SG (1994) Introduction of YACs into intact yeast cells by a procedure which shows low levels of recombinagenicity and co-transformation. Nucleic Acids Res 22, 5011–5015
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
Schedl A, Larin Z, Montoliu L, Thies E, Kelsey G, Lehrach H, Schutz G (1993) A method for the generation of YAC transgenic mice by pro-nuclear microinjection. Nucleic Acids Res 21, 4783–4787
Smith DR, Smythe AP, Moir DT (1990) Amplification of large artificial chromosomes. Proc Natl Acad Sci USA 87, 8242–8246
Tsui LC (1992) The spectrum of cystic fibrosis mutations. Trends Genet 8, 392–398
Zehetner G, Lehrach H (1994) The reference library system—sharing biological material and experimental data. Nature 367, 498–491
Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM (1994) Positional cloning of the mouse obese gene and its human homologue. Nature 372, 425–432
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Toye, A.A., Schalkwyk, L., Lehrach, H. et al. A yeast artificial chromosome (YAC) library containing 10 haploid chicken genome equivalents. Mammalian Genome 8, 274–276 (1997). https://doi.org/10.1007/s003359900408
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s003359900408