Yeast artificial chromosomes

From OpenWetWare

Revision as of 18:45, 2 January 2007 by Tk (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search

Yeast artificial chromosomes (YACs) are synthetic double stranded linear constructs containing the elements necessary for replication as independent chromosomes in yeast. These elements are:

  • an autonomous replication sequence (ARS): ARS1, chromosome III ARS, ARSH4
  • a centromere: CEN4, CEN6: centromeres consist of three centromere determining elements, CDE I, CDE II, and CDE III.
  • a telomeric sequence at each end

Typically the chromosome also contains a selection marker such as TRP1, Lys2 or Ura3.

Minimal size for a YAC is between 50kb and 100kb, while maximum sizes are 1Mb to 3Mb.

A common tool for constructing YACs is a shuttle plasmid such as pYAC4 which replicates in E. coli, has a multiple cloning site, and a pair of telomeres which can be cleaved to form a linear fragment. Available as an E.coli plasmid ATCC 67379, sequence at U01086.

There are two common centromere sequences, CEN4 and CEN6. CEN4 is found in most yeast centromere-containiing vectors, such as pYAC4. These vectors typically use ARS1 sequences.

The pRS313- pRS316 plasmids use the CEN6 + ARSH4 cassette (Sikorski89).

Yeast strains

  • AB1380 (MATa ura3-52 trp1 lys2-1 ade2-1 can1-100 his5) (Burke87) ATCC 204682
  • J57D (MATa, ura3-52, trp1 ade2-101 can1-100 leu2-3, 112 his3-6) (Haldi96)
    • claimed to be 2-3x more efficient at large insert cloning than AB1380
  • W303-1a (MATa leu2-3,112 trp1-1 can1-100 ura3-1 ade2-1 his3-11,15 ybp1-1) Rothstein notes (see Veal03) Open Biosystems: YSC1058
  • YPH500 (MATα ura3-52 lys2-801_amber ade2-101_ochre trp1-Δ63 his1-Δ200 leu2-Δ1) ATCC:204680 (Sikorski89)
  • S288C (MATα SUC2 gal2 mal mel flo1 flo8-1 hap1) ATCC: 204508 (Mortimer86)

YAC Plasmids

  • pYAC4 (Burke87) has Sup4 for ade2 selection, interrupted by the cloning site (EcoRI), ura3, a pair of facing telomeres surrounding a his3 gene (linearized and cut out with BamHI), a pBR322 based E. coli replication and ampR region, a barely functional TRP1 gene, ARS1 and CEN4. Sequence at Genbank U01086
  • pJS97 / pJS98 plasmids (Shero91) are available at ATCC 77191 as a two-plasmid kit. Each plasmid supplies one chromosome end.
    • pJS97 has CEN4 and ARSH4, URA3 gene, Sup11 tRNA for ade2 selection, a pUC19 derived E. coli replication region, including ampR, and a telomere.
    • pJS98 has ARSH4, a functional TRP1 gene, a pUC19 derived E. coli replication region including ampR, and a telomere.
  • pCGS966 (Smith90, Smith92, Moir93) has ARS1 on both arms, Gal inducible extra copy production, NeoR for mammalian expression and a functional promoter for the Trp gene, unlike pYAC4.


  1. Hieter P, Pridmore D, Hegemann JH, Thomas M, Davis RW, and Philippsen P. . pmid:2996783. PubMed HubMed [Hieter85]
  2. Mann C and Davis RW. . pmid:3537685. PubMed HubMed [Mann86]
  3. Mortimer RK and Johnston JR. . pmid:3519363. PubMed HubMed [Moritmer86]
  4. Burke DT, Carle GF, and Olson MV. . pmid:3033825. PubMed HubMed [Burke87]
  5. Cottarel G, Shero JH, Hieter P, and Hegemann JH. . pmid:2552293. PubMed HubMed [Cottarel89]
  6. Anand R, Villasante A, and Tyler-Smith C. . pmid:2542900. PubMed HubMed [Anand89]
  7. Sikorski RS and Hieter P. . pmid:2659436. PubMed HubMed [Sikorski89]
  8. Sgaramella V, Ferretti L, Damiani G, and Sora S. . pmid:2189412. PubMed HubMed [Sgaramella90]
  9. Smith DR, Smyth AP, and Moir DT. . pmid:2236036. PubMed HubMed [Smith90]
  10. Wells RA, Germino GG, Krishna S, Buckle VJ, and Reeders ST. . pmid:2276741. PubMed HubMed [Wells90]
  11. Van Houten JV and Newlon CS. . pmid:2196439. PubMed HubMed [VanHouten90]
  12. McCormick MK, Antonarakis SE, and Hieter P. . pmid:2091693. PubMed HubMed [McCormick90]
  13. Burke DT. . pmid:2091697. PubMed HubMed [Burke90]
  14. Shero JH, McCormick MK, Antonarakis SE, and Hieter P. . pmid:2071158. PubMed HubMed [Shero92]
  15. Burke DT and Olson MV. . pmid:2005791. PubMed HubMed [Burke91]
  16. Connelly C, McCormick MK, Shero J, and Hieter P. . pmid:2045094. PubMed HubMed [Connelly91]
  17. Smith DR, Smyth AP, and Moir DT. . pmid:1336106. PubMed HubMed [Smith92]
  18. Ragoussis J, Trowsdale J, and Markie D. . pmid:1620611. PubMed HubMed [Ragoussis92]
  19. de Bruin D, Lanzer M, and Ravetch JV. . pmid:1427849. PubMed HubMed [deBruin92]
  20. Deshpande AM and Newlon CS. . pmid:1406623. PubMed HubMed [Deshpande92]
  21. Marahrens Y and Stillman B. . pmid:1536007. PubMed HubMed [Marahrens92]
  22. Christianson TW, Sikorski RS, Dante M, Shero JH, and Hieter P. . pmid:1544568. PubMed HubMed [Christianson92]
  23. Moir DT, Dorman TE, Smyth AP, and Smith DR. . pmid:8462878. PubMed HubMed [Moir93]
  24. Spencer F, Ketner G, Connelly C, and Hieter P. Targeted recombination-based cloning and manipulation of large DNA segments in yeast. Methods: A companion to methods in enzymology 1993; 5:161-175 (no pubmed entry)


  25. Kuhn RM and Ludwig RA. . pmid:8163163. PubMed HubMed [Kuhn94]
  26. Hugerat Y, Spencer F, Zenvirth D, and Simchen G. . pmid:7959756. PubMed HubMed [Hugerat94]
  27. Spencer F, Hugerat Y, Simchen G, Hurko O, Connelly C, and Hieter P. . pmid:7959757. PubMed HubMed [Spencer94]
  28. Hamer L, Johnston M, and Green ED. . pmid:8524833. PubMed HubMed [Hamer95]
  29. Mahmood A, Kimura T, Takenaka M, and Yoshida K. . pmid:8841544. PubMed HubMed [Mahmood95]
  30. Mahmood A, Kimura T, Takenaka M, and Yoshida K. . pmid:8880145. PubMed HubMed [Mahmood96]
  31. Haldi ML, Strickland C, Lim P, VanBerkel V, Chen X, Noya D, Korenberg JR, Husain Z, Miller J, and Lander ES. . pmid:8854865. PubMed HubMed [Haldi96]
  32. US Patent 5776745 Ketner et al. 1998


  33. Kouprina N, Nikolaishvili N, Graves J, Koriabine M, Resnick MA, and Larionov V. . pmid:10087193. PubMed HubMed [Kooprina99]
  34. Cocchia M, Kouprina N, Kim SJ, Larionov V, Schlessinger D, and Nagaraja R. . pmid:10954614. PubMed HubMed [Cocchia00]
  35. Veal EA, Ross SJ, Malakasi P, Peacock E, and Morgan BA. . pmid:12743123. PubMed HubMed [Veal03]
All Medline abstracts: PubMed HubMed
Personal tools