Molecular analysis of Saccharomyces cerevisiae chromosome I : On the number of genes and the identification of essential genes using temperature-sensitive-lethal mutations
dc.contributor.author | Harris, Steven D. | en_US |
dc.contributor.author | Cheng, Judy | en_US |
dc.contributor.author | Pugh, Tom A. | en_US |
dc.contributor.author | Pringle, John R. | en_US |
dc.date.accessioned | 2006-04-10T15:13:33Z | |
dc.date.available | 2006-04-10T15:13:33Z | |
dc.date.issued | 1992-05-05 | en_US |
dc.identifier.citation | Harris, Steven D., Cheng, Judy, Pugh, Tom A., Pringle, John R. (1992/05/05)."Molecular analysis of Saccharomyces cerevisiae chromosome I : On the number of genes and the identification of essential genes using temperature-sensitive-lethal mutations." Journal of Molecular Biology 225(1): 53-65. <http://hdl.handle.net/2027.42/30054> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6WK7-4DMP5N2-12/2/3ca38729e8e9877beed4982808b1284f | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/30054 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=1583694&dopt=citation | en_US |
dc.description.abstract | Previous analyses of Saccharomyces cerevisiae chromosome I have suggested that the majority (>75%) of single-copy essential genes on this chromosome are difficult or impossible to identify using temperature-sensitive (Ts-) lethal mutations. To investigate whether this situation reflects intrinsic difficulties in generating temperature-sensitive proteins or constraints on mutagenesis in yeast, we subjected three cloned essential genes from chromosome I to mutagenesis in an Escherichia coli mutator strain and screened for Ts- lethal mutations in yeast using the "plasmid-shuffle" technique. We failed to obtain Ts- lethal mutations in two of the genes (FUN12 and FUN20), while the third gene yielded such mutations, but only at a low frequency. DNA sequence analysis of these mutant alleles and of the corresponding wild-type region revealed that each mutation was a single substitution not in the previously identified gene FUN19, but in the adjacent, newly identified essential gene FUN53. FUN19 itself proved to be non-essential. These results suggest that many essential proteins encoded by genes on chromosome I cannot be rendered thermolabile by single mutations. However, the results obtained with FUN53 suggest that there may also be significant constraints on mutagenesis in yeast. The 5046 base-pair interval sequenced contains the complete FUN19, FUN53 and FUN20 coding regions, as well as a portion of the adjacent non-essential FUN21 coding region. In all, 68 to 75% of this interval is open reading frame. None of the four predicted products shows significant homologies to known proteins in the available databases. | en_US |
dc.format.extent | 1957497 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | Molecular analysis of Saccharomyces cerevisiae chromosome I : On the number of genes and the identification of essential genes using temperature-sensitive-lethal mutations | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Natural Resources and Environment | en_US |
dc.subject.hlbsecondlevel | Molecular, Cellular and Developmental Biology | en_US |
dc.subject.hlbsecondlevel | Ecology and Evolutionary Biology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Biology The University of Michigan, Ann Arbor, MI 48109-1048, USA | en_US |
dc.contributor.affiliationum | Department of Biology The University of Michigan, Ann Arbor, MI 48109-1048, USA | en_US |
dc.contributor.affiliationum | Department of Biology The University of Michigan, Ann Arbor, MI 48109-1048, USA | en_US |
dc.contributor.affiliationum | Department of Biology The University of Michigan, Ann Arbor, MI 48109-1048, USA | en_US |
dc.identifier.pmid | 1583694 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/30054/1/0000422.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1016/0022-2836(92)91025-K | en_US |
dc.identifier.source | Journal of Molecular Biology | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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