A yeast artificial chromosome (YAC) is a vector used to clone DNA fragments larger than 100 kb and up to 3000 kb. YACs are useful for the physical mapping of complex genomes and for the cloning of large genes. First described in 1983 by Murray and Szostak, a YAC is an artificially constructed chromosome that contains a centromere, telomeres and an autonomous replicating sequence (ARS) element, which are required for replication and preservation of YAC in yeast cells. ARS elements are thought to act as replication origins. A YAC is built using an initial circular plasmid, which is typically broken into two linear molecules using restriction enzymes; DNA ligase is then used to ligate a sequence or gene of interest between the two linear molecules, forming a single large linear piece of DNA..
A plasmid-derived origin of replication (ori) and an antibiotic resistance gene allow the YAC vector to be amplified and selected for in E.coli. TRP1 and URA3 genes are included in the YAC vector to provide a selection system for identifying transformed yeast cells that include YAC by complementing recessive alleles trp1 and ura3 in yeast host cell. YAC vector cloning site for foreign DNA is located within the SUP4 gene. This gene compensates for a mutation in the yeast host cell that causes the accumulation of red pigment. The host cells are normally red, and those transformed with YAC only, will form colourless colonies. Cloning of a foreign DNA fragment into the YAC causes insertional inactivation, restoring the red colour. Therefore the colonies that contain the foreign DNA fragment are red.
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Famous quotes containing the words yeast, artificial and/or chromosome:
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—Carmen McRae (b. 1922)
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—Alexander Herzen (1812–1870)
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—Frank Pittman (20th century)