Abstract
Soon after the first report of how yeast artificial chromosomes (YACs) could be used as cloning vectors for large DNA fragments, the transfer of YACs into mammalian cells came into focus of interest, Following mammalian cell transfer, the YAC integrates into the host genome. Because of the large size of YACs, genes contained within the construct should be regulated and expressed at levels comparable to their endogenous counterparts. This method should, therefore, allow the identification of genes by complementation and can also be used to study gene function and regulation in vivo. YACs ranging in size from 35–650 kb have been transferred to mammalian cells.
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Notes
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Note added in proofs. The thymidine kinase gene (TK), present in amplification vectors to apply strong selective pressure, contains a cryptic promoter in its coding sequence, which leads to expression in the testes. Recent results have shown that this can confer male sterility in transgenic mice (2). Establishment of transgenic lines from YAC constructs carrying amplification vectors might, therefore, be difficult.
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© 1996 Humana Press Inc.
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Schedl, A., Grimes, B., Montoliu, L. (1996). YAC Transfer by Microinjection. In: Markie, D. (eds) YAC Protocols. Methods in Molecular Biology™, vol 54. Humana Press. https://doi.org/10.1385/0-89603-313-9:293
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DOI: https://doi.org/10.1385/0-89603-313-9:293
Publisher Name: Humana Press
Print ISBN: 978-0-89603-313-9
Online ISBN: 978-1-59259-541-9
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