Yeast artificial chromosomes: Difference between revisions
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The pRS313- pRS316 plasmids use the CEN6 + ARSH4 cassette (Sikorski89). | The pRS313- pRS316 plasmids use the CEN6 + ARSH4 cassette (Sikorski89). | ||
The pCGS966 plasmid (Smith90, Smith92, Moir93) has ARS1 on both arms, Gal inducible extra copy production, NeoR for mammalian expression | |||
Minimal size for a YAC is between 50kb and 100kb, while maximum sizes are 1Mb to 3Mb. | Minimal size for a YAC is between 50kb and 100kb, while maximum sizes are 1Mb to 3Mb. | ||
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# Sikorski89 pmid=2659436 | # Sikorski89 pmid=2659436 | ||
# Sgaramella90 pmid=2189412 | # Sgaramella90 pmid=2189412 | ||
# Smith90 pmid=2236036 | |||
# Wells90 pmid=2276741 | # Wells90 pmid=2276741 | ||
# VanHouten90 pmid=2196439 | # VanHouten90 pmid=2196439 |
Revision as of 18:32, 3 August 2006
Yeast artificial chromosomes (YACs) are synthetic double stranded linear constructs containing the elements necessary for replication in yeast. These elements are:
- an autonomous replication sequence (ARS): ARS1, chromosome III ARS, ARSH4
- a centromere: CEN4
- a telomeric sequence at each end
Typically the chromosome also contains a selection marker such as Lys2 or Ura3.
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. Yeast host AB1380 is available as ATCC 204682.
The pRS313- pRS316 plasmids use the CEN6 + ARSH4 cassette (Sikorski89).
The pCGS966 plasmid (Smith90, Smith92, Moir93) has ARS1 on both arms, Gal inducible extra copy production, NeoR for mammalian expression
Minimal size for a YAC is between 50kb and 100kb, while maximum sizes are 1Mb to 3Mb.
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