Restriction mapping of yeast artificial chromosomes by UV-mediated partial digestion

1994 ◽  
Vol 5 (9) ◽  
pp. 588-590 ◽  
Author(s):  
C. Nobile ◽  
A. Forabosco
Keyword(s):  
Restriction Mapping ◽  
Artificial Chromosomes ◽  
Partial Digestion ◽  
Yeast Artificial Chromosomes

Arabidopsis YAC restriction mapping

Genome ◽  
1998 ◽  
Vol 41 (6) ◽  
pp. 806-817
Author(s):  
Shaun M Morroll ◽  
Zoe A Wilson
Keyword(s):  
Positional Cloning ◽  
Floral Development ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Genomic Region ◽  
Restriction Mapping ◽  
Artificial Chromosomes ◽  
Number Of Genes ◽  
Yeast Artificial Chromosomes ◽  
Large Genomic Region

The approach of partial restriction mapping and vector hybridisation has been used to restriction map and align six yeast artificial chromosomes (YACs) corresponding to the top arm (~27.9 centiMorgans, cM) of Arabidopsis chromosome 5 and confirm the chimeric nature of a further four clones which map to this region. The restriction endonucleases Sma1 and Sfi1 which recognise rare-medium frequency sites in the Arabidopsis genome were used. This work has restriction mapped a 315 kb region that includes a number of genes implicated in floral development, namely PISTILLATA and TOUSLED, and a number of uncharacterised genes involved in male gametogenesis (e.g., Ms1 and Ms37). The information generated can be used to transcriptionally map genes to this contig and will provide data for the isolation of several uncharacterised floral development genes which lie in this region. This approach has demonstrated how large tracts of YAC DNA can be mapped and aligned to show the presence/absence of chimeric YAC clones and provide detailed restriction knowledge for a large genomic region to help facilitate the positional cloning of genes.Key words: yeast artificial chromosome, YAC, Arabidopsis thaliana, partial restriction mapping, floral development.

scholarly journals Yeast artificial chromosomes: an alternative approach to the molecular analysis of mouse developmental mutations

1990 ◽  
Vol 56 (2-3) ◽  
pp. 203-208 ◽  
Author(s):  
Zoia Larin ◽  
Hans Lehrach
Keyword(s):  
Mouse Genome ◽  
Yeast Artificial Chromosome ◽  
Germ Line ◽  
Artificial Chromosome ◽  
Molecular Study ◽  
Insert Size ◽  
Artificial Chromosomes ◽  
Partial Digestion ◽  
Genomic Regions ◽  
Yeast Artificial Chromosomes

SummaryMammalian genetics now allows a molecular study of genomic regions previously analysed by genetic and embryological techniques. To simplify such an analysis, we have established a number of libraries of mouse DNA in Yeast Artificial Chromosome (YAC) vectors, constructed either by partial digestion with EcoRI, or by complete digestion with enzymes which cut rarely in the mammalian genome. In this paper we report the construction of complete digest libraries prepared from mouse genomic DNA using the rare cutter enzymes NoiI and BssHII, and the detection of gene loci from the H-2 complex, the t–complex, and other loci from the mouse genome. Due to their large insert size, YAC clones simplify the cloning of extended regions of the mouse genome surrounding known developmental mutations and should, after introduction into the germ line, offer a high probability of correct expression of the genes contained within the cloned region. We hope that this will allow the use of YAC clones to scan regions of interest such as the t–complex for specific genes by testing DNA introduced into transgenic mice for the ability to complement mutations localised to this region.

scholarly journals Cellular gels. Purifying and mapping long DNA molecules

1991 ◽  
Vol 273 (3) ◽  
pp. 695-699 ◽  
Author(s):  
P H Dear ◽  
P R Cook
Keyword(s):  
Baker’S Yeast ◽  
Restriction Mapping ◽  
Baker's Yeast ◽  
Dna Molecules ◽  
Chromosomal Dna ◽  
Artificial Chromosomes ◽  
Chromosome Iii ◽  
Yeast Artificial Chromosomes ◽  
Pulsed Fields ◽  
Protective Coat

Long DNA molecules of greater than 10(5) bp (0.1 Mbp) are easily broken by pipetting. Therefore, chromosomal DNA is generally isolated after embedding cells in a protective coat of agarose. The embedded DNA can then be cut into long pieces and fractionated on gels using pulsed fields, but these pieces are again easily broken if the resolved DNA molecules are recovered from the gels. We now describe a novel gel matrix, a ‘cellular’ gel, that permits the recovery of resolved fragments from gels in a form that enables facile manipulation without shear. This facilitates purification and restriction mapping of fragments of 0.1-1.0 Mbp. We illustrate the utility of the method by mapping chromosome III of baker's yeast, which has a length of approximately 0.36 Mbp. This method should facilitate purification and restriction mapping of yeast artificial chromosomes.

Murine Albino-Deletion Complex: High-Resolution Microsatellite Map and Genetically Anchored YAC Framework Map

Genetics ◽  
1997 ◽  
Vol 147 (2) ◽  
pp. 787-799
Author(s):  
Brad A Rikke ◽  
Dabney K Johnson ◽  
Thomas E Johnson
Keyword(s):  
Positional Cloning ◽  
Interspecific Backcross ◽  
Genetic Distances ◽  
Sequence Length ◽  
Recombination Rates ◽  
Oak Ridge ◽  
Artificial Chromosomes ◽  
Microsatellite Map ◽  
Specific Locus ◽  
Yeast Artificial Chromosomes

The murine albino-deletion complex developed as part of the Oak Ridge specific-locus test covers 6–11 cM of chromosome 7. This complex has proven to be a valuable resource for localizing traits to a small target region suitable for positional cloning. In this study, we mapped the endpoints of deletions in this complex using all of the available Mit simple-sequence length polymorphism (SSLP) markers. Concurrently, this mapping has determined the map order of nearly all of the SSLP markers, most of which were previously unresolved. The SSLP-based deletion map was confirmed and genetic distances were determined using the European Collaborative Interspecific Backcross panel of nearly a thousand mice. The average SSLP marker resolution is 0.3–0.4 cM, comparable to the cloning capacity of yeast artificial chromosomes (YACs). The SSLP markers were then used to construct a genetically anchored YAC framework map that further confirms the deletion map. We find that the largest deleted region distal to Tyr is about two to three times larger than the largest proximal deleted region, and the original C3H/101 regions flanking the deletions (moved to an St2A cch/cch background) are smaller than anticipated, which we suggest may result from increased recombination rates immediately flanking the deleted regions.

A Novel Ty1-Mediated Fragmentation Method for Native and Artificial Yeast Chromosomes Reveals That the Mouse Steel Gene is a Hotspot for Ty1 Integration

Genetics ◽  
1996 ◽  
Vol 143 (2) ◽  
pp. 673-683
Author(s):  
Jacob Z Dalgaard ◽  
Mukti Banerjee ◽  
M Joan Curcio
Keyword(s):  
Transcription Unit ◽  
Yeast Genome ◽  
Physical Analysis ◽  
Single Chromosome ◽  
Artificial Chromosomes ◽  
Unique Site ◽  
High Fraction ◽  
A Site ◽  
Yeast Artificial Chromosomes ◽  
Random Insertion

Abstract We have developed a powerful new tool for the physical analysis of genomes called Ty1-mediated chromosomal fragmentation and have used the method to map 24 retrotransposon insertions into two different mousederived yeast artificial chromosomes (YACs). Expression of a plasmid-encoded GAL1:Ty1 fusion element marked with the retrotransposition indicator gene, ade2AI, resulted in a high fraction of cells that sustained a single Ty1 insertion marked with ADE2. Strains in which Ty1ADE2 inserted into aYAC were identified by cosegregation of the ADE2 gene with the URA3-marked YAC. Ty1ADE2 elements also carried a site for the endonuclease I-DmoI, which we demonstrate is not present anywhere in the yeast genome. Consequently, I-DmoI cleaved a single chromosome or YAC at the unique site of Ty1ADE2 insertion, allowing rapid mapping of integration events. Our analyses showed that the frequency of Ty1ADE2 integration into YACs is equivalent to or higher than that expected based on random insertion. Remarkably, the 50-kb transcription unit of the mouse Steel locus was shown to be a highly significant hotspot for Ty1 integration. The accessibility of mammalian transcription units to Ty1 insertion stands in contrast to that of yeast transcription units.

Construction and Characterization of a Partial Library of Yeast Artificial Chromosomes from Human Chromosome 21

1991 ◽  
Vol 10 (4) ◽  
pp. 301-310 ◽  
Author(s):  
M. BELLIS ◽  
A. GÉRARD ◽  
J.P. CHARLIEU ◽  
B. MARÇAIS ◽  
M.E. BRUN ◽  
...  
Keyword(s):  
Human Chromosome ◽  
Chromosome 21 ◽  
Artificial Chromosomes ◽  
Human Chromosome 21 ◽  
Yeast Artificial Chromosomes

scholarly journals Detection of homologous recombination between yeast artificial chromosomes with overlapping inserts

1991 ◽  
Vol 19 (5) ◽  
pp. 997-1000 ◽  
Author(s):  
Alessandra Cellini ◽  
Rosa M. Lacatena ◽  
Glauco p. tocchini-Valentini
Keyword(s):  
Homologous Recombination ◽  
Artificial Chromosomes ◽  
Yeast Artificial Chromosomes

Complete coverage of the Schizosaccharomyces pombe genome in yeast artificial chromosomes

1992 ◽  
Vol 1 (4) ◽  
pp. 273-277 ◽  
Author(s):  
Elmar Maier ◽  
Jörg D. Hoheisel ◽  
Linda McCarthy ◽  
Richard Mott ◽  
Andrei V. Grigoriev ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Complete Coverage ◽  
Artificial Chromosomes ◽  
Yeast Artificial Chromosomes
Sign in / Sign up

Export Citation Format

Share Document