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Yeast Artificial Chromosomes

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Molecular Biomethods Handbook

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Abstract

Since their mtroductlon in 1987, yeast artificial chromosomes (commonly referred to as YACs) have had a tremendous impact on the field of molecular genetics The ability of YACs to carry and propagate DNA fragments of up to -2 mllhon base pairs in size has been instrumental in the construction of the first- and second-generation physical maps of the human genome (1,2) YACs have assisted in the generation of detailed maps of discrete chromosomal regions and slgmficantly speeded up posltlonal cloning projects leading to the ldentlficatlon and cloning of a variety of genes involved in inherited diseases (3,4) YACs have also been used for studies on gene function via complementatlon of mutations as well as for the transfer of complete genomlc gene sequences for the generation of transgemc animals Widespread use of this broad range of appllcatlons has been facilitated by the avallabrhty of a number of high-quality genomlc YAC libraries via centralized screening centers This clearly illustrates the scientific advances that can be made when tools and technology are shared in the spirit of cooperation.

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References

  1. Chumakov, I, Welssenbach, J, and Cohen, D (1993) A first generation physical map of the human genome Nature 336,698–701

    Google Scholar 

  2. Chumakov, I, Rlgault, P, LeGall, I, and Cohen, D (1995) A YAC contig map of the human genome Nature 377(Suppl.), Genome Dir 175–297

    Google Scholar 

  3. Collins, F S (1992) Posltlonal cloning Lets not call it reverse anymore Nat Genet 1,3–6

    Article  PubMed  CAS  Google Scholar 

  4. Nelson, D L (1995) Posltlonal cloning comes of age Curr Opznzon Gen Dev 5,298–303

    Article  CAS  Google Scholar 

  5. Herskowltz, I (1988) Life cycle of the budding yeast Saccharomyces cerevzslae Mwroblol Rev 52, 536

    Google Scholar 

  6. Struhl, K, Stmchomb, D T, Scherer, S, and Davis, R W (1979) High frequency transformation of yeast Autonomous replication of hybrid DNA molecules Proc Nati Acad Sci USA 76, 1035–1039

    Article  CAS  Google Scholar 

  7. Stmchomb, D T, Struhl, K, and Davis, R W (1979) Isolation and characterization of a yeast chromosomal replicator Nature 282,39–43

    Article  Google Scholar 

  8. Bouton, A H and Smith, M in (1986) Fine-structure analysis of the DNA sequence requirement for autonomous rephcatlon of Saccharomyces cerevlslae plasmids Mol Cell Blol 6,2354–2357

    CAS  Google Scholar 

  9. Clarke, L and Carbon, J (1980) Isolation of a yeast centromere and construction of functional small circular chromosomes Nature 287, 504–509

    Article  PubMed  CAS  Google Scholar 

  10. Szostak, J W and Blackbum, E H (1982) Cloning yeast telomeres on linear plasmld vectors Cell 29,245–255

    Article  PubMed  CAS  Google Scholar 

  11. Murray, A W and Szostak, J W (1983) Construction of artificial chromosomes in yeast Nature 305, 189–193

    Article  PubMed  CAS  Google Scholar 

  12. Burke, D T, Carie, G F, and Olson, M V (1987) Clomng of large segments of exogenous DNA into yeast by means of artificial chromosome vectors Science 236,806–812

    Article  PubMed  CAS  Google Scholar 

  13. Burke, D T and Olson, M V (1991) Preparation of clone libraries in yeast artlticlal chromsosome vectors Meth Enzymol 194,251–270

    Article  PubMed  CAS  Google Scholar 

  14. Bartholdl, M F, Meyne, J, Albright, K, Luedemann, M, Campbell, E, Chntton, D, Daven, L, Van Dllla, M, and Cram, L S (1987) Chromosome sorting by flow cytometry, in Methods in Enzymology voi 15 1 (Gottesman M,ed ), Academic, San Diego, CA, pp252–267

    Google Scholar 

  15. Abbott, C and Povey, S (1994) Somatzc Cell Hybrzds The Basics (Beynond, R, Brown, T A, and Howe, C,eds ), Oxford University Press, Cambridge, UK

    Google Scholar 

  16. Abide, F E, Wada, M, Little, R D, and Schlessinger, D (1990) Yeast artificial chromosomes containing human Xq2-Xq28 DNA Library construction and representation of probe sequences Genomics 7,363–376

    Article  Google Scholar 

  17. Den Dunnen, J T, Grootscholten, P in, Dauwerse, J G, Walker, A P, Monaco, A P, Butler, R, Anand, R, Coffey, A J, Bentley, D. R, Steensma, H Y, and van Ommen, G J B (1992) Reconstructdlon of the 2 4 Mb human DMD-gene by homologous YAC recombmatlon Hum Mol Genet 1, 19–23.

    Google Scholar 

  18. McCormick, M K, Campbell, E, Deaven, L, and Moyzls, R K (1993) Low frequency chlmerlc yeast artificial chromosome libraries from flow-sorted human chromosomes 16 and 21 Proc Nati Acad Sci USA 90,1063–1067

    Article  CAS  Google Scholar 

  19. McCormick, M K, Buckler, A, Bruno, W, Campbell, E, Shera, K, Tomey, D, Deaven, L, and Moyzls, R (1993) Construction and charactensatlon of a YAC library with a low frequency of chunenc clones from flow-sorted human chromosome 9 Genomics 18,553–558

    Article  PubMed  CAS  Google Scholar 

  20. Little, R D, Porta, G, Carle, G F, Schlessinger, D., and D’Urso, M (1989) Yeast artlticlal chromosomes with 200 to 800 kllobase msrts of human DNA containing HLA, Vk, 5s and Xq24-Xq28 sequences Proc Nati Acad Sci USA 86,1598–1602

    Article  CAS  Google Scholar 

  21. Edwards, K J, Thompson, H, Edwards, D, deSalzleu1, A, Sparks, C, Thompson, S A, Greenland, A J, Eyeres, M, and Schuch, W (1992) Construction and characterlsatlon of a yeast artlficlal chromosome library containing three haplold maize genome equivalents Plant Mol Biol 19,299–308

    Article  CAS  Google Scholar 

  22. Coulson, A R, Waterston, R, Klff, J, Sulston, J, and Kohara, Y (1988) Genome Imking with yeast artificial chromosomes Nature 335, 181–186

    Article  Google Scholar 

  23. Chartier, F L, Keer, J T, Sutchffe, M J., Hennques, D A., Mlleham, P, and Brown, S D in (1992) Construction of a mouse yeast artificial chromosome library in a recombmatlon-deficient strain of yeast Nat Genet 1, 132–136

    Article  PubMed  CAS  Google Scholar 

  24. Brownstem, 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 chromosomes Science 244, 1348–1351

    Article  Google Scholar 

  25. 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, charactensatlon, screening and storage Nucleic Acids Res 18, 1951–1956

    Article  PubMed  CAS  Google Scholar 

  26. Larm, Z, Monaco, A P, and Lehrach, H (1991) Yeast artificial chromosome libraries contaming large inserts from mouse and human DNA Proc Nati Acad Sci USA 88, 4123–4127

    Article  Google Scholar 

  27. Albertsen, H in, Abderrahlm, H, Cann, H in, Dausset, J, LePasher, D, and Cohen, D (1990) ConstructIon and charactelsatlon of a yeast arnficlal chromosome library contaming seven haplold human genome equivalents Proc Nati Acad Sci USA 87,4256–4260

    Article  CAS  Google Scholar 

  28. Jones, M H, KhwaJa, 0 S A, Bnggs, H, Lambson, B, Davey, P in, Chalmers, J, Zhou, C-Y, Walker, E in, Zhang, Y, Todd, C, Ferguson-Smith, M A, and Affara, N A (1994) A set of ninety-seven overlapping yeast artificial chromosome clones spanning the human Y chromosome euchromatm Genomics 24,266–275

    Article  PubMed  CAS  Google Scholar 

  29. Anand, R (1995) Cloning into yeast artlficlal chromosomes, in DNA Clonzng 3 A Practzcal Approach (Glover, D in and Hames, B D,eds ), Oxford Umverslty Press, Cambridge, UK, pp 103–125

    Google Scholar 

  30. Ross, M T, Hohelsel, J D, Monaco, A P, Larm, Z, Zehetner, G, and Lehrach, H (1992) High-density gadded YAC filters their potential as genome mapping tools, in Technzquesfor the Analyszs of Complex Genomes (Anand, R,ed ), Academic, San Diego, CA, pp 137–153

    Google Scholar 

  31. Bentley, D R, Todd, C, Collms, J, Holland, J, Dunham, I, Hassock, S, Bankler, A, and Glanelh, F (1992) The development and appllcatlon of automated ridding for efficlent screening of yeast and bactertal ordered libraries Genomics 12,534–541

    Article  PubMed  CAS  Google Scholar 

  32. Schwartz, D C and Cantor, C R (1984) Separatton of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresls Cell 37, 67–75

    Article  PubMed  CAS  Google Scholar 

  33. Lal, E, Barren, B W, Clark, S in, Simon, M I, and Hood, L (1989) Pulsed field gel electrophoresls Blotechnrques 7,34–42

    Google Scholar 

  34. Monaco, A. P (1995) Pulsed Field Gel Electrophoresls A Practical Approach(Rlckwood, D and Hames, B D,eds ), Oxford Umverslty Press, Cambndge, UK

    Google Scholar 

  35. Trask, B J (1991) Fluorescence in situ hybrldlsatlon applications in cytogenetlcs and gene mapping Trend Genet 7, 149–154

    CAS  Google Scholar 

  36. Buckle, V J and Rack, K A (1993) Fluorescent in situ hybndlsatlon, in Human Genetzc Dzsease Analyszs A Pructzcal Approach (Davies, K E, ed ), Oxford Umverslty Press, Cambridge, UK, pp 59–82

    Google Scholar 

  37. Silverman, G A (1993) Isolating vector-insert Junctions from yeast artlficlal chromosomes PCR Meth Appl 3, 141–150

    CAS  Google Scholar 

  38. Sliverman, G A, Ye, R D, Pollock, K in, 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 Nati Acad Sci USA 86,7485–7489

    Article  Google Scholar 

  39. Nelson, D L, Ballablo, A, Victoria, M F, Plerettl, M, Bles, R D, Gibbs, R A, Maley, J A, Chmault A C, Webster, T D,and Caskey, C T (1991) Mu-primed polymerase chain reaction for regional assignment of 110 yeast artificial chromosome clones from the human X chromosome Identlficatron of clones associated with a disease locus Proc Nati Acad Sci USA 88,6157–6161

    Article  CAS  Google Scholar 

  40. Riley, J, Butler, R, Ogllvle, D, Fmmear, R, Jenner, D, Powell, S, Anand, R, Smith, J C, and Markham, A F (1990) A novel, rapid method for the lsolatlon of terminal sequences from yeast artlficlal chromosome (YAC) clones Nuclezc Aczds Res 18, 2887–2890

    Article  CAS  Google Scholar 

  41. Weber, J L (1990) Abundant class of human DNA polymorphisms which can be typed using the polymerase cham reactlon Genomlcs 7,524–530

    Article  CAS  Google Scholar 

  42. Cornells, F, Hashlmoto, L, Lovendge, J, MacCarthy, A, Buckle, V, Juller, C, and Bell, J (1992) Identlficatlon of a CA repeat at the TCRA locus using yeast artlficlal chromosomes A general method for generating highly polymorphic markers at chosen loci Genomzcs 13,820–825

    Article  Google Scholar 

  43. Ilzuka, M, Makm, R, Seklya, T, and Hayashl, K (1993) Selective lsolatlon of,hlghly polymorphic (dC-dA)n-(dG-dT)n mlcrosatelhtes by stringent hybndlsatlon GATA 10,2–5

    Google Scholar 

  44. Brown, W R A and Bird, A P (1986) Long-range restrlctlon site mapping of mammalian genomlc DNA Nature 322,477–481

    Article  PubMed  CAS  Google Scholar 

  45. Bird, A P (1987) CpG islands as gene markers in the vertebrate nucleus Trends Genet 3,342–346

    Article  CAS  Google Scholar 

  46. Elvm, P, Butler, R, and Hedge, P J (1992) Transcribed sequences wlthm YACs HTF island cloning and cDNA library screening, in Techniques of the Analyszs of Complex Genomes (Anand, R, ed ), Academic, San Diego, CA, pp 155–171

    Google Scholar 

  47. Sealey, P G, Whlttaker, P A, and Southern, E in (1985) Removal of repeated sequences from hybndlsatlon probes Nucleic Acids Res 13, 1905–1932

    Article  PubMed  CAS  Google Scholar 

  48. Panmoo, S, PatanJah, S R, Shukla, H, Chaplin, D D., and Weissman, S in (1991) cDNA selection Efficient PCR approach for the selection of cDNAs encoded in large chromosomal DNA fragments Proc Nati Acad Sci USA 88,9623–9627

    Article  Google Scholar 

  49. Lovett, M, Kere, J, and Hinton, L in (1991) Direct selection a method for the lsolatlon of large genomlc regions Proc Nati Acad Sci USA 88,9628–9632

    Article  CAS  Google Scholar 

  50. Auch, D and Reth, M (1990) Exon trap cloning Using PCR to rapidly detect and clone exons from genomtc DNA fragments Nucleic Acids Res 25,6743,6744

    Google Scholar 

  51. Duyk, G in, Kim, S, Myers, R in, and Cox, D R (1990) Exon trapping A genettc screen to identify candidate transcribed sequences in cloned mammahan DNA Proc Nati Acad Sci USA 87,8995–8999

    Article  CAS  Google Scholar 

  52. Buckler, A J, Chang, D D, Graw, S L, Brook, J D., Haber, D A, Sharp, P A, and Housman, D E (1991) Exon amphfication A strategy to isolate mammalian genes based on RNA splicing Proc Nati Acad Sci USA 88,40054009

    Article  Google Scholar 

  53. Gmrke, A, Barnes, T S, Patterson, D., Schrld, D, Featherstone, T, and Olson, M V (1991) Clomng and in vtvo expression of the human GART gene using yeast arttfitciarll chromosomes EMBO J 10, 1629–1634

    Google Scholar 

  54. Gmrke, A and Huxley, C (1991) Transfer of the human HPRT and GART genes from yeast to mamimahan cells by mrrcromJectton of YAC DNA Som Cell Mol Genet 17, 573–580

    Google Scholar 

  55. Pachms, V, Pevuey, L, Rothstem, R, and Constanttm, F (1990) Transfer of a yeast artifictal chromosome carrying human DNA from Saccharomyces cerevzszae into mammalian cells Proc Nati Acad Sci USA 87,5109–5113

    Article  Google Scholar 

  56. Pavan, W J, Hteter, P, and Reeves, R H (1990) Modtficatton and transfer into an embryonal carcmoma cell line of a 360-krlobase human-derived yeast artificial chromosome Mol Cell Blol 10,4163–4169

    CAS  Google Scholar 

  57. Snvastava, A K and Schlessinger, D (1991) Vectors for inserting selectable markers in vector arms and human DNA inserts of yeast arttfictal chromosomes (YACs) Gene 103,53059

    Google Scholar 

  58. Riley, J H, Morten, J E N, and Anand, R (1992) Targeted integration of neomycm into yeast artificial chromosomes (YACs) for transfectton into mammahan cells Nuclezc Acids Res 20,2971–2976

    Article  CAS  Google Scholar 

  59. Forget, B G (1993) YAC transgenes bigger is probably better (Review) Proc Nati Acad Sci USA 90,7909–7911

    Article  CAS  Google Scholar 

  60. Peterson, K R., Clegg, C H, Huxley, C, Josephson, B in, Haugen, H S, Furukawa, T, and Stamatoyannopoulos, G (1993) Transgemc mice containing a 248-kb yeast arttfictal chromosome carrying the human beta-globm locus display proper developmental control of human globm genes Proc Nati Acad Sci USA 90,7593–7597

    Article  CAS  Google Scholar 

  61. Frazer, K A, Narla, G, Zhang, J L, and Rubm, E in (1995) The apohpoprotem(a) gene 1s regulated by sex hormones, and acute-phase inducers in YAC transgemc Nat Gen 9,424–431

    Article  CAS  Google Scholar 

  62. Huxley, C, Passage, E, Manson, A, Putzu, G, Ftgarella-Branger, D, Pelhssier, J F, and Fontes, M (1996) Constructton of a mouse model of Charcot-Mane-Tooth disease type 1A by pronuclear inJectton of human YAC DNA Hum Mol Genet 5,563–569

    Article  PubMed  CAS  Google Scholar 

  63. Sternberg, N (1990) Bacteriophage PI cloning system for the tsolation, ampltfication, and recovery of DNA fragments as large as 100 ktlobase pairs Proc Nati Acad Scz USA 87,103–107

    Article  CAS  Google Scholar 

  64. Ioannou, P A, et al (1994) A new bacteriophage Pl-derived vector for the propagation of large human DNA fragments Nat Genet 6,84–89

    Article  PubMed  CAS  Google Scholar 

  65. Shizuya, H, et al (1992)Cloning and stable maintenance of 30-ktlobase-pan fragments of human DNA in Eschrertchta cold using an F-factor-based vector Proc Natl Acad Scr USA 89,8794–8797

    Article  CAS  Google Scholar 

  66. Huxley, C (1994) Mammalian artifictal chromosomes a new tool for gene therapy Gene Ther 1,7–12

    PubMed  CAS  Google Scholar 

  67. Harrington, J J, Van Bokkelen, G, Mays, R W, Gustashaw, K, and Willard, H F (1997) Formatton of de novo centromeres and construction of first-generation human arttfictal mtcrochromosomes Nature Genet 15,345–355

    Article  PubMed  CAS  Google Scholar 

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Flannery, A., Anand, R. (1998). Yeast Artificial Chromosomes. In: Rapley, R., Walker, J.M. (eds) Molecular Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-59259-642-3_24

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  • DOI: https://doi.org/10.1007/978-1-59259-642-3_24

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-501-0

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