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Journal articles on the topic 'Artificial Yeast Chromosomes'

Author: Grafiati

Published: 4 June 2021

Last updated: 16 February 2022

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1

Ross, L. O., D. Treco, A. Nicolas, J. W. Szostak, and D. Dawson. "Meiotic recombination on artificial chromosomes in yeast." Genetics 131, no. 3 (July 1, 1992): 541–50. http://dx.doi.org/10.1093/genetics/131.3.541.

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Abstract We have examined the meiotic recombination characteristics of artificial chromosomes in Saccharomyces cerevisiae. Our experiments were carried out using minichromosome derivatives of yeast chromosome III and yeast artificial chromosomes composed primarily of bacteriophage lambda DNA. Tetrad analysis revealed that the artificial chromosomes exhibit very low levels of meiotic recombination. However, when a 12.5-kbp fragment from yeast chromosome VIII was inserted into the right arm of the artificial chromosome, recombination within that arm mimicked the recombination characteristics of the fragment in its natural context including the ability of crossovers to ensure meiotic disjunction. Both crossing over and gene conversion (within the ARG4 gene contained within the fragment) were measured in the experiments. Similarly, a 55-kbp region from chromosome III carried on a minichromosome showed crossover behavior indistinguishable from that seen when it is carried on chromosome III. We discuss the notion that, in yeast, meiotic recombination behavior is determined locally by small chromosomal regions that function free of the influence of the chromosome as a whole.

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2

Blackburn, Elizabeth H. "Artificial chromosomes in yeast." Trends in Genetics 1 (January 1985): 8–12. http://dx.doi.org/10.1016/0168-9525(85)90007-1.

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3

Sambrook, Joseph, and David W. Russell. "Working with Yeast Artificial Chromosomes." Cold Spring Harbor Protocols 2006, no. 1 (June 2006): pdb.prot3297. http://dx.doi.org/10.1101/pdb.prot3297.

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4

Selleri, Licia, Gary G. Hermanson, James H. Eubanks, and Glen A. Evans. "Chromosomal in situ hybridization using yeast artificial chromosomes." Genetic Analysis: Biomolecular Engineering 8, no. 2 (April 1991): 59–66. http://dx.doi.org/10.1016/1050-3862(91)90050-2.

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5

Pavan, W. J., and R. H. Reeves. "Integrative selection of human chromosome-specific yeast artificial chromosomes." Proceedings of the National Academy of Sciences 88, no. 17 (September 1, 1991): 7788–91. http://dx.doi.org/10.1073/pnas.88.17.7788.

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6

McCormick, M. K., J. H. Shero, M. C. Cheung, Y. W. Kan, P. A. Hieter, and S. E. Antonarakis. "Construction of human chromosome 21-specific yeast artificial chromosomes." Proceedings of the National Academy of Sciences 86, no. 24 (December 1, 1989): 9991–95. http://dx.doi.org/10.1073/pnas.86.24.9991.

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7

Coulson, Alan, Robert Waterston, Jane Kiff, John Sulston, and Yuji Kohara. "Genome linking with yeast artificial chromosomes." Nature 335, no. 6186 (September 1988): 184–86. http://dx.doi.org/10.1038/335184a0.

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8

Ragoussis, Jiannis, John Trowsdale, and David Markie. "Mitotic recombination of yeast artificial chromosomes." Nucleic Acids Research 20, no. 12 (1992): 3135–38. http://dx.doi.org/10.1093/nar/20.12.3135.

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9

Cai, H., P. Kiefel, J. Yee, and I. Duncan. "A yeast artificial chromosome clone map of the Drosophila genome." Genetics 136, no. 4 (April 1, 1994): 1385–99. http://dx.doi.org/10.1093/genetics/136.4.1385.

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Abstract We describe the mapping of 979 randomly selected large yeast artificial chromosome (YAC) clones of Drosophila DNA by in situ hybridization to polytene chromosomes. Eight hundred and fifty-five of the clones are euchromatic and have primary hybridization sites in the banded portions of the polytene chromosomes, whereas 124 are heterochromatic and label the chromocenter. The average euchromatic clone contains about 211 kb and, at its primary site, labels eight or nine contiguous polytene bands. Thus, the extent as well as chromosomal position of each clone has been determined. By direct band counts, we estimate our clones provide about 76% coverage of the euchromatin of the major autosomes, and 63% coverage of the X. When previously reported YAC mapping data are combined with ours, euchromatic coverage is extended to about 90% for the autosomes and 82% for the X. The distribution of gap sizes in our map and the coverage achieved are in good agreement with expectations based on the assumption of random coverage, indicating that euchromatic clones are essentially randomly distributed. However, certain gaps in coverage, including the entire fourth chromosome euchromatin, may be significant. Heterochromatic sequences are underrepresented among the YAC clones by two to three fold. This may result, at least in part, from underrepresentation of heterochromatic sequences in adult DNA (the source of most of the clones analyzed), or from clone instability.

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10

Izvolsky, K. "Yeast artificial chromosome segregation from host chromosomes with similar lengths." Nucleic Acids Research 26, no. 21 (November 1, 1998): 5011–12. http://dx.doi.org/10.1093/nar/26.21.5011.

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11

Zhong, Tao P., Kim Kaphingst, Uma Akella, Maryann Haldi, Eric S. Lander, and Mark C. Fishman. "Zebrafish Genomic Library in Yeast Artificial Chromosomes." Genomics 48, no. 1 (February 1998): 136–38. http://dx.doi.org/10.1006/geno.1997.5155.

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12

Larin, Z. "A method for linking yeast artificial chromosomes." Nucleic Acids Research 24, no. 21 (November 1, 1996): 4192–96. http://dx.doi.org/10.1093/nar/24.21.4192.

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13

Huxley, Clare, and Andreas Gnirke. "Transfer of yeast artificial chromosomes from yeast to mammalian cells." BioEssays 13, no. 10 (October 1991): 545–50. http://dx.doi.org/10.1002/bies.950131009.

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14

Wang, Z. X. "Physical Mapping of Rice Chromosome 1 with Yeast Artificial Chromosomes (YACs)." DNA Research 3, no. 5 (January 1, 1996): 291–96. http://dx.doi.org/10.1093/dnares/3.5.291.

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15

Poorkaj, Parvoneh, Kenneth R. Peterson, and Gerard D. Schellenberg. "Single-Step Conversion of P1 and P1 Artificial Chromosome Clones into Yeast Artificial Chromosomes." Genomics 68, no. 1 (August 2000): 106–10. http://dx.doi.org/10.1006/geno.2000.6267.

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16

Murray, A. W., and J. W. Szostak. "Construction and behavior of circularly permuted and telocentric chromosomes in Saccharomyces cerevisiae." Molecular and Cellular Biology 6, no. 9 (September 1986): 3166–72. http://dx.doi.org/10.1128/mcb.6.9.3166.

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We developed techniques that allow us to construct novel variants of Saccharomyces cerevisiae chromosomes. These modified chromosomes have precisely determined structures. A metacentric derivative of chromosome III which lacks the telomere-associated X and Y' elements, which are found at the telomeres of most yeast chromosomes, behaves normally in both mitosis and meiosis. We made a circularly permuted telocentric version of yeast chromosome III whose closest telomere was 33 kilobases from the centromere. This telocentric chromosome was lost at a frequency of 1.6 X 10(-5) per cell compared with a frequency of 4.0 X 10(-6) for the natural metacentric version of chromosome III. An extremely telocentric chromosome whose closet telomere was only 3.5 kilobases from the centromere was lost at a frequency of 6.0 X 10(-5). The mitotic stability of telocentric chromosomes shows that the very high frequency of nondisjunction observed for short linear artificial chromosomes is not due to inadequate centromere-telomere separation.

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17

Murray, A. W., and J. W. Szostak. "Construction and behavior of circularly permuted and telocentric chromosomes in Saccharomyces cerevisiae." Molecular and Cellular Biology 6, no. 9 (September 1986): 3166–72. http://dx.doi.org/10.1128/mcb.6.9.3166-3172.1986.

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We developed techniques that allow us to construct novel variants of Saccharomyces cerevisiae chromosomes. These modified chromosomes have precisely determined structures. A metacentric derivative of chromosome III which lacks the telomere-associated X and Y' elements, which are found at the telomeres of most yeast chromosomes, behaves normally in both mitosis and meiosis. We made a circularly permuted telocentric version of yeast chromosome III whose closest telomere was 33 kilobases from the centromere. This telocentric chromosome was lost at a frequency of 1.6 X 10(-5) per cell compared with a frequency of 4.0 X 10(-6) for the natural metacentric version of chromosome III. An extremely telocentric chromosome whose closet telomere was only 3.5 kilobases from the centromere was lost at a frequency of 6.0 X 10(-5). The mitotic stability of telocentric chromosomes shows that the very high frequency of nondisjunction observed for short linear artificial chromosomes is not due to inadequate centromere-telomere separation.

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18

Saji, Shoko, Yosuke Umehara, Baltazar A. Antonio, Hiroko Yamane, Hiroshi Tanoue, Tomoya Baba, Hiroyoshi Aoki, et al. "A physical map with yeast artificial chromosome (YAC) clones covering 63% of the 12 rice chromosomes." Genome 44, no. 1 (February 1, 2001): 32–37. http://dx.doi.org/10.1139/g00-076.

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A new YAC (yeast artificial chromosome) physical map of the 12 rice chromosomes was constructed utilizing the latest molecular linkage map. The 1439 DNA markers on the rice genetic map selected a total of 1892 YACs from a YAC library. A total of 675 distinct YACs were assigned to specific chromosomal locations. In all chromosomes, 297 YAC contigs and 142 YAC islands were formed. The total physical length of these contigs and islands was estimated to 270 Mb which corresponds to approximately 63% of the entire rice genome (430 Mb). Because the physical length of each YAC contig has been measured, we could then estimate the physical distance between genetic markers more precisely than previously. In the course of constructing the new physical map, the DNA markers mapped at 0.0-cM intervals were ordered accurately and the presence of potentially duplicated regions among the chromosomes was detected. The physical map combined with the genetic map will form the basis for elucidation of the rice genome structure, map-based cloning of agronomically important genes, and genome sequencing.Key words: physical mapping, YAC contig, rice genome, rice chromosomes.

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19

Jinks-Robertson, Sue, Shariq Sayeed, and Tamara Murphy. "Meiotic Crossing Over Between Nonhomologous Chromosomes Affects Chromosome Segregation in Yeast." Genetics 146, no. 1 (May 1, 1997): 69–78. http://dx.doi.org/10.1093/genetics/146.1.69.

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Meiotic recombination between artificial repeats positioned on nonhomologous chromosomes occurs efficiently in the yeast Saccharomyces cerevisiae. Both gene conversion and crossover eventS have been observed, with crossovers yielding reciprocal translocations. In the current study, 5.5-kb ura3 repeats positioned on chromosomes V and XV were used to examine the effect of ectopic recombination on meiotic chromosome segregation. Ura+ random spores were selected and gene conversion vs. crossover events were distinguished by Southern blot analysis. Approximately 15% of the crossover events between chromosomes V and XV were associated with missegregation of one of these chromosomes. The missegregation was manifest as hyperploid spores containing either both translocations plus a normal chromosome, or both normal chromosomes plus one of the translocations. In those cases where it could be analyzed, missegregation occurred at the first meiotic division. These data are discussed in terms of a model in which ectopic crossovers compete efficiently with normal allelic crossovers in directing meiotic chromosome segregation.

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20

van Brabant, Anja J., Walton L. Fangman, and Bonita J. Brewer. "Active Role of a Human Genomic Insert in Replication of a Yeast Artificial Chromosome." Molecular and Cellular Biology 19, no. 6 (June 1, 1999): 4231–40. http://dx.doi.org/10.1128/mcb.19.6.4231.

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ABSTRACT Yeast artificial chromosomes (YACs) are a common tool for cloning eukaryotic DNA. The manner by which large pieces of foreign DNA are assimilated by yeast cells into a functional chromosome is poorly understood, as is the reason why some of them are stably maintained and some are not. We examined the replication of a stable YAC containing a 240-kb insert of DNA from the human T-cell receptor beta locus. The human insert contains multiple sites that serve as origins of replication. The activity of these origins appears to require the yeast ARS consensus sequence and, as with yeast origins, additional flanking sequences. In addition, the origins in the human insert exhibit a spacing, a range of activation efficiencies, and a variation in times of activation during S phase similar to those found for normal yeast chromosomes. We propose that an appropriate combination of replication origin density, activation times, and initiation efficiencies is necessary for the successful maintenance of YAC inserts.

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21

Silverman, G. A. "Isolating vector-insert junctions from yeast artificial chromosomes." Genome Research 3, no. 3 (December 1, 1993): 141–50. http://dx.doi.org/10.1101/gr.3.3.141.

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22

Peterson, Kenneth R., Christopher H. Clegg, and Qiliang Li. "Production of transgenic mice with yeast artificial chromosomes." Trends in Genetics 13, no. 2 (February 1997): 61–66. http://dx.doi.org/10.1016/s0168-9525(97)01003-2.

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23

Montoliu, L., A. Schedl, G. Kelsey, P. Lichter, Z. Larin, H. Lehrach, and G. Schutz. "Generation of Transgenic Mice with Yeast Artificial Chromosomes." Cold Spring Harbor Symposia on Quantitative Biology 58 (January 1, 1993): 55–62. http://dx.doi.org/10.1101/sqb.1993.058.01.009.

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24

Garza, D., J. Ajioka, D. Burke, and D. Hartl. "Mapping the Drosophila genome with yeast artificial chromosomes." Science 246, no. 4930 (November 3, 1989): 641–46. http://dx.doi.org/10.1126/science.2510296.

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25

Davies, Nicholas P., and Marianne Brüggemann. "Extension of yeast artificial chromosomes by cosmid multimers." Nucleic Acids Research 21, no. 3 (1993): 767–68. http://dx.doi.org/10.1093/nar/21.3.767.

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26

Ross, Lyle O., Drora Zenvirth, Ana Rita Jardim, and Dean Dawson. "Double-strand breaks on artificial chromosomes in yeast." Chromosoma 109, no. 4 (July 5, 2000): 226–34. http://dx.doi.org/10.1007/s004129900049.

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27

Sutcliffe, James S., Fuping Zhang, C. Thomas Caskey, David L. Nelson, and Stephen T. Warren. "PCR amplification and analysis of yeast artificial chromosomes." Genomics 13, no. 4 (August 1992): 1303–6. http://dx.doi.org/10.1016/0888-7543(92)90051-s.

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28

Infante, A., S. Lo, and J. L. Hall. "A Chlamydomonas genomic library in yeast artificial chromosomes." Genetics 141, no. 1 (September 1, 1995): 87–93. http://dx.doi.org/10.1093/genetics/141.1.87.

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Abstract We have constructed and characterized a Chlamydomonas reinhardtii total genomic library in yeast artificial chromosomes (YACs). The library contains 7500 clones with inserts ranging in size from 100-200 kb. The representation of the library was assessed by screening one-third of it with a probe derived from the dispersed repeat, Gulliver, which occurs approximately 13 times in the genome. At least 10 of these Gulliver loci were isolated within 15 independent YACs. Two of these YACs encompass the Gulliver element designated G, which was reported to map to the uni linkage group (ULG). The end clones of these two YACs have been genetically mapped by RFLP analysis in an interspecific cross and thereby shown to be closely linked to the APM locus on the ULG. A third uni-specific YAC has also been isolated and its ends have been mapped by RFLP analysis. Genetic and RFLP analysis of these and other YACs indicates that the frequency of chimeric YACs in the library is very low. The library was constructed in a second generation vector that enables plasmid rescue of YAC end clones as well as copy number amplification of artificial chromosomes. We provide evidence that amplification of intact YACs requires a rad1:rad52 yeast strain.

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29

Palmer, R. E., E. Hogan, and D. Koshland. "Mitotic transmission of artificial chromosomes in cdc mutants of the yeast, Saccharomyces cerevisiae." Genetics 125, no. 4 (August 1, 1990): 763–74. http://dx.doi.org/10.1093/genetics/125.4.763.

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Abstract In the yeast, Saccharomyces cerevisiae, cell division cycle (CDC) genes have been identified whose products are required for the execution of different steps in the cell cycle. In this study, the fidelity of transmission of a 14-kb circular minichromosome and a 155-kb linear chromosome fragment was examined in cell divisions where specific CDC products were temporarily inactivated with either inhibitors, or temperature sensitive mutations in the appropriate CDC gene. All of the cdc mutants previously shown to induce loss of endogenous linear chromosomes also induced loss of a circular minichromosome and a large linear chromosome fragment in our study (either 1:0 or 2:0 loss events). Therefore, the efficient transmission of these artificial chromosomes depends upon the same trans factors that are required for the efficient transmission of endogenous chromosomes. In a subset of cdc mutants (cdc6, cdc7 and cdc16), the rate of minichromosome loss was significantly greater than the rate of loss of the linear chromosome fragment, suggesting that a structural feature of the minichromosome (nucleotide content, length or topology) makes the minichromosome hypersensitive to the level of function of these CDC gene products. In another subset of cdc mutants (cdc7 and cdc17), the relative rate of 1:0 events to 2:0 events differed for the minichromosome and chromosome fragment, suggesting that the type of chromosome loss event observed in these mutants was dependent upon chromosome structure. Finally, we show that 2:0 events for the minichromosome can occur by both a RAD52 dependent and RAD52 independent mechanism. These results are discussed in the context of the molecular functions of the CDC products.

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30

Sato, Hiroshi, and Shigeaki Saitoh. "Switching the centromeres on and off: epigenetic chromatin alterations provide plasticity in centromere activity stabilizing aberrant dicentric chromosomes." Biochemical Society Transactions 41, no. 6 (November 20, 2013): 1648–53. http://dx.doi.org/10.1042/bst20130136.

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The kinetochore, which forms on a specific chromosomal locus called the centromere, mediates interactions between the chromosome and the spindle during mitosis and meiosis. Abnormal chromosome rearrangements and/or neocentromere formation can cause the presence of multiple centromeres on a single chromosome, which results in chromosome breakage or cell cycle arrest. Analyses of artificial dicentric chromosomes suggested that the activity of the centromere is regulated epigenetically; on some stably maintained dicentric chromosomes, one of the centromeres no longer functions as a platform for kinetochore formation, although the DNA sequence remains intact. Such epigenetic centromere inactivation occurs in cells of various eukaryotes harbouring ‘regional centromeres’, such as those of maize, fission yeast and humans, suggesting that the position of the active centromere is determined by epigenetic markers on a chromosome rather than the nucleotide sequence. Our recent findings in fission yeast revealed that epigenetic centromere inactivation consists of two steps: disassembly of the kinetochore initiates inactivation and subsequent heterochromatinization prevents revival of the inactivated centromere. Kinetochore disassembly followed by heterochromatinization is also observed in normal senescent human cells. Thus epigenetic centromere inactivation may not only stabilize abnormally generated dicentric chromosomes, but also be part of an intrinsic mechanism regulating cell proliferation.

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31

Chang, E., J. Luna, J. Giacalone, D. Uyar, GA Silverman, and U. Francke. "Regional localization of 56 new human chromosome 18-specific yeast artificial chromosomes." Cytogenetic and Genome Research 65, no. 1-2 (1994): 136–39. http://dx.doi.org/10.1159/000133620.

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32

Stock, W., S. C. Chandrasekharappa, W. L. Neuman, M. M. LeBeau, B. H. Brownstein, and C. A. Westbrook. "Characterization of yeast artificial chromosomes containing interleukin genes on human chromosome 5." Cytogenetic and Genome Research 61, no. 4 (1992): 263–65. http://dx.doi.org/10.1159/000133417.

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33

Scherer, Stephen W., Brock J. F. Tompkins, and Lap-Chee Tsui. "A human Chromosome 7-specific genomic DNA library in yeast artificial chromosomes." Mammalian Genome 3, no. 3 (1992): 179–81. http://dx.doi.org/10.1007/bf00352464.

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34

Guacci, V., E. Hogan, and D. Koshland. "Chromosome condensation and sister chromatid pairing in budding yeast." Journal of Cell Biology 125, no. 3 (May 1, 1994): 517–30. http://dx.doi.org/10.1083/jcb.125.3.517.

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We have developed a fluorescent in situ hybridization (FISH) method to examine the structure of both natural chromosomes and small artificial chromosomes during the mitotic cycle of budding yeast. Our results suggest that the pairing of sister chromatids: (a) occurs near the centromere and at multiple places along the chromosome arm as has been observed in other eukaryotic cells; (b) is maintained in the absence of catenation between sister DNA molecules; and (c) is independent of large blocks of repetitive DNA commonly associated with heterochromatin. Condensation of a unique region of chromosome XVI and the highly repetitive ribosomal DNA (rDNA) cluster from chromosome XII were also examined in budding yeast. Interphase chromosomes were condensed 80-fold relative to B form DNA, similar to what has been observed in other eukaryotes, suggesting that the structure of interphase chromosomes may be conserved among eukaryotes. While additional condensation of budding yeast chromosomes were observed during mitosis, the level of condensation was less than that observed for human mitotic chromosomes. At most stages of the cell cycle, both unique and repetitive sequences were either condensed or decondensed. However, in cells arrested in late mitosis (M) by a cdc15 mutation, the unique DNA appeared decondensed while the repetitive rDNA region appeared condensed, suggesting that the condensation state of separate regions of the genome may be regulated differently. The ability to monitor the pairing and condensation of sister chromatids in budding yeast should facilitate the molecular analysis of these processes as well as provide two new landmarks for evaluating the function of important cell cycle regulators like p34 kinases and cyclins. Finally our FISH method provides a new tool to analyze centromeres, telomeres, and gene expression in budding yeast.

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35

Krizman, D. B., T. A. Hofmann, U. DeSilva, E. D. Green, P. S. Meltzer, and J. M. Trent. "Identification of 3'-terminal exons from yeast artificial chromosomes." Genome Research 4, no. 6 (June 1, 1995): 322–26. http://dx.doi.org/10.1101/gr.4.6.322.

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36

Sheehan, Catherine, and Anthony S. Weiss. "Yeast artificial chromosomes: rapid extraction for high resolution analysis." Nucleic Acids Research 18, no. 8 (1990): 2193. http://dx.doi.org/10.1093/nar/18.8.2193.

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37

Schlessinger, David, and Ramaiah Nagaraja. "Impact and implications of yeast and human artificial chromosomes." Annals of Medicine 30, no. 2 (January 1998): 186–91. http://dx.doi.org/10.3109/07853899808999402.

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38

Springer, P. S., K. J. Edwards, and J. L. Bennetzen. "DNA class organization on maize Adh1 yeast artificial chromosomes." Proceedings of the National Academy of Sciences 91, no. 3 (February 1, 1994): 863–67. http://dx.doi.org/10.1073/pnas.91.3.863.

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39

Di Rienzo, Anna, Amy Peterson, Soma Das, and Nelson B. Freimer. "Genome mapping by arbitrary amplification of yeast artificial chromosomes." Mammalian Genome 4, no. 7 (July 1993): 359–63. http://dx.doi.org/10.1007/bf00360585.

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40

Korenberg, J. R., X. N. Chen, S. Mitchell, S. Fannin, S. Gerwehr, D. Cohen, and I. Chumakov. "A high-fidelity physical map of human chromosome 21q in yeast artificial chromosomes." Genome Research 5, no. 5 (December 1, 1995): 427–43. http://dx.doi.org/10.1101/gr.5.5.427.

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41

Wang, S. Y. "A High-Resolution Physical Map of Human Chromosome 21p Using Yeast Artificial Chromosomes." Genome Research 9, no. 11 (November 1, 1999): 1059–73. http://dx.doi.org/10.1101/gr.9.11.1059.

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42

Hermanson, Gary G., Merl F. Hoekstra, David L. McElligott, and Glen A. Evans. "Rescue of end fragments of yeast artificial chromosomes by homologous recombination in yeast." Nucleic Acids Research 19, no. 18 (1991): 4943–48. http://dx.doi.org/10.1093/nar/19.18.4943.

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43

Hamer, L., M. Johnston, and E. D. Green. "Isolation of yeast artificial chromosomes free of endogenous yeast chromosomes: construction of alternate hosts with defined karyotypic alterations." Proceedings of the National Academy of Sciences 92, no. 25 (December 5, 1995): 11706–10. http://dx.doi.org/10.1073/pnas.92.25.11706.

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44

Brown, W. R., M. J. Dobson, and P. MacKinnon. "Telomere cloning and mammalian chromosome analysis." Journal of Cell Science 95, no. 4 (April 1, 1990): 521–26. http://dx.doi.org/10.1242/jcs.95.4.521.

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Although eucaryotic chromosomes vary in size over five orders of magnitude and are constituents of diverse genetic systems the fundamental features of their telomeres appear to be almost completely conserved. This can be exploited to enable molecular cloning of human telomeres in yeast and suggests that many of the ideas that will arise from studies of telomeres in the experimentally tractable ciliates and yeasts will hold true of mammalian telomeres. The particular value of cloned mammalian telomeres is that they contribute reagents for mapping mammalian chromosomes and that they provide one set of elements for the construction of artificial mammalian chromosomes.

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45

Weise, Anja, Peter Harbarth, Uwe Claussen, and Thomas Liehr. "Fluorescence In Situ Hybridization (FISH) on Human Chromosomes Using Photoprobe Biotin-labeled Probes." Journal of Histochemistry & Cytochemistry 51, no. 4 (April 2003): 549–51. http://dx.doi.org/10.1177/002215540305100418.

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Fluorescence in situ hybridization (FISH) on human chromosomes in meta-and interphase is a well-established technique in clinical and tumor cytogenetics and for studies of evolution and interphase architecture. Many different protocols for labeling the DNA probes used for FISH have been published. Here we describe for the first time the successful use of Photoprobe biotin-labeled DNA probes in FISH experiments. Yeast artificial chromosome (YAC) and whole chromosome painting (wcp) probes were tested.

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46

Schmidt, Dagmar, Marion S. Röder, Harald Dargatz, Norbert Wolf, Günther F. Schweizer, Andy Tekauz, and Martin W. Ganal. "Construction of a YAC library from barley cultivar Franka and identification of YAC-derived markers linked to the Rh2 gene conferring resistance to scald (Rhynchosporium secalis)." Genome 44, no. 6 (December 1, 2001): 1031–40. http://dx.doi.org/10.1139/g01-108.

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The Rh2 resistance gene of barley (Hordeum vulgare) confers resistance against the scald pathogen (Rhynchosporium secalis). A high-resolution genetic map of the Rh2 region on chromosome 1 (7H) was established by the use of molecular markers. Tightly linked markers from this region were used to screen existing and a newly constructed yeast artificial chromosome (YAC) library of barley cv. Franka composed of 45 000 clones representing approximately two genome equivalents. Corresponding YAC clones were identified for most markers, indicating that the combined YAC library has good representation of the barley genome. The contiguous sets of YAC clones with the most tightly linked molecular markers represent entry points for map-based cloning of this resistance gene.Key words: yeast artificial chromosomes, map-based cloning, disease resistance gene, library screening, Hordeum vulgare.

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47

Montoliu, L., A. Schedl, G. Kelsey, H. Zentgraf, P. Lichter, and G. Schutz. "Germ line transmission of yeast artificial chromosomes in transgenic mice." Reproduction, Fertility and Development 6, no. 5 (1994): 577. http://dx.doi.org/10.1071/rd9940577.

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Several groups have recently reported the successful generation of transgenic mice harbouring yeast artificial chromosomes (YACs). Different methodological approaches have been shown to produce similar results, namely, the faithful expression of the transgenes carried on YAC DNA. In this paper, we compare the reported techniques for obtaining transgenic mice carrying YACs using a 250-kb YAC bearing the mouse tyrosinase gene. These methods include: microinjection of gel-purified YAC DNA into pronuclei of fertilized mouse oocytes, yeast spheroblast fusion with embryonic stem (ES) cells and lipofection of YAC DNA into ES cells. Taken together, these reports show that the delivery of large genomic regions covering a gene of interest (such as those cloned in YAC vectors) is feasible, and will ensure appropriate temporal and spatial expression of the transgene at a level comparable to that of the endogenous counterpart.

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Chumakov, I. M., I. Le Gall, A. Billault, P. Ougen, P. Soularue, S. Guillou, P. Rigault, et al. "Isolation of chromosome 21–specific yeast artificial chromosomes from a total human genome library." Nature Genetics 1, no. 3 (June 1992): 222–25. http://dx.doi.org/10.1038/ng0692-222.

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Fuchs, Jörg, Dorothee-U. Kloos, Martin W. Ganal, and Ingo Schubert. "In situ localization of yeast artificial chromosome sequences on tomato and potato metaphase chromosomes." Chromosome Research 4, no. 4 (June 1996): 277–81. http://dx.doi.org/10.1007/bf02263677.

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Simpson, K., A. McGuigan, and C. Huxley. "Stable episomal maintenance of yeast artificial chromosomes in human cells." Molecular and Cellular Biology 16, no. 9 (September 1996): 5117–26. http://dx.doi.org/10.1128/mcb.16.9.5117.

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Abstract:

Plasmids carrying the Epstein-Barr virus origin of plasmid replication (oriP) have been shown to replicate autonomously in latently infected human cells (J. Yates, N. Warren, D. Reisman, and B. Sugden, Proc. Natl. Acad. Sci. USA 81:3806-3810, 1984). We demonstrate that addition of this domain is sufficient for stable episomal maintenance of yeast artificial chromosomes (YACs), up to at least 660 kb, in human cells expressing the viral protein EBNA-1. To better approximate the latent viral genome, YACs were circularized before addition of the oriP domain by homologous recombination in yeast cells. The resulting OriPYACs were maintained as extrachromosomal molecules over long periods in selection; a 90-kb OriPYAC was unrearranged in all cell lines analyzed, whereas the intact form of a 660-kb molecule was present in two of three cell lines. The molecules were also relatively stable in the absence of selection. This finding indicates that the oriP-EBNA-1 interaction is sufficient to stabilize episomal molecules of at least 660 kb and that such elements do not undergo rearrangements over time. Fluorescence in situ hybridization analysis demonstrated a close association of OriPYACs, some of which were visible as pairs, with host cell chromosomes, suggesting that the episomes replicate once per cell cycle and that stability is achieved by attachment to host chromosomes, as suggested for the viral genome. The wide availability of YAC libraries, the ease of manipulation of cloned sequences in yeast cells, and the episomal stability make OriPYACs ideal for studying gene function and control of gene expression.

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