Incorporation of copy-number control elements into yeast artificial chromosomes by targeted homologous recombination

1993 ◽  
Vol 4 (3) ◽  
pp. 141-147 ◽  
Author(s):  
Douglas R. Smith ◽  
Adrienne P. Smyth ◽  
William M. Strauss ◽  
Donald T. Moir
Keyword(s):  
Homologous Recombination ◽  
Copy Number ◽  
Artificial Chromosomes ◽  
Copy Number Control ◽  
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

scholarly journals A Chlamydomonas genomic library in yeast artificial chromosomes.

Genetics ◽  
1995 ◽  
Vol 141 (1) ◽  
pp. 87-93
Author(s):  
A Infante ◽  
S Lo ◽  
J L Hall
Keyword(s):  
Linkage Group ◽  
Copy Number ◽  
Yeast Strain ◽  
Genomic Library ◽  
Interspecific Cross ◽  
Rflp Analysis ◽  
Plasmid Rescue ◽  
Artificial Chromosomes ◽  
Yeast Artificial Chromosomes ◽  
Copy Number Amplification

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.

scholarly journals Rescue of end fragments of yeast artificial chromosomes by homologous recombination in yeast

1991 ◽  
Vol 19 (18) ◽  
pp. 4943-4948 ◽  
Author(s):  
Gary G. Hermanson ◽  
Merl F. Hoekstra ◽  
David L. McElligott ◽  
Glen A. Evans
Keyword(s):  
Homologous Recombination ◽  
Artificial Chromosomes ◽  
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.

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