scholarly journals Meiotic recombination on artificial chromosomes in yeast.

Genetics ◽  
1992 ◽  
Vol 131 (3) ◽  
pp. 541-550 ◽  
Author(s):  
L O Ross ◽  
D Treco ◽  
A Nicolas ◽  
J W Szostak ◽  
D Dawson
Keyword(s):  
Meiotic Recombination ◽  
Artificial Chromosome ◽  
Artificial Chromosomes ◽  
Crossover Behavior ◽  
Yeast Chromosome ◽  
Low Levels ◽  
Chromosome Iii ◽  
The Right ◽  
Yeast Artificial Chromosomes ◽  
Derivatives Of

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.

scholarly journals Involvement of Sir2/4 in Silencing of DNA Breakage and Recombination on Mouse YACs during Yeast Meiosis

2005 ◽  
Vol 16 (3) ◽  
pp. 1449-1455 ◽  
Author(s):  
Yair Klieger ◽  
Ofer Yizhar ◽  
Drora Zenvirth ◽  
Neta Shtepel-Milman ◽  
Margriet Snoek ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Hot Spot ◽  
Yeast Cells ◽  
Class Iii ◽  
Dna Double Strand Breaks ◽  
Artificial Chromosomes ◽  
Mouse Major Histocompatibility Complex ◽  
Dna Backbone ◽  
Yeast Meiosis ◽  
Yeast Artificial Chromosomes

Yeast artificial chromosomes (YACs) that contain human DNA backbone undergo DNA double-strand breaks (DSBs) and recombination during yeast meiosis at rates similar to the yeast native chromosomes. Surprisingly, YACs containing DNA covering a recombination hot spot in the mouse major histocompatibility complex class III region do not show meiotic DSBs and undergo meiotic recombination at reduced levels. Moreover, segregation of these YACs during meiosis is seriously compromised. In meiotic yeast cells carrying the mutations sir2 or sir4, but not sir3, these YACs show DSBs, suggesting that a unique chromatin structure of the YACs, involving Sir2 and Sir4, protects the YACs from the meiotic recombination machinery. We speculate that the paucity of DSBs and recombination events on these YACs during yeast meiosis may reflect the refractory nature of the corresponding region in the mouse genome.

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.

Single-Step Conversion of P1 and P1 Artificial Chromosome Clones into Yeast Artificial Chromosomes

Genomics ◽  
2000 ◽  
Vol 68 (1) ◽  
pp. 106-110 ◽  
Author(s):  
Parvoneh Poorkaj ◽  
Kenneth R. Peterson ◽  
Gerard D. Schellenberg
Keyword(s):  
Artificial Chromosome ◽  
Single Step ◽  
Artificial Chromosomes ◽  
Yeast Artificial Chromosomes

scholarly journals Use of a Recombination Reporter Insert To Define Meiotic Recombination Domains on Chromosome III ofSaccharomyces cerevisiae

1999 ◽  
Vol 19 (7) ◽  
pp. 4832-4842 ◽  
Author(s):  
Valérie Borde ◽  
Tzu-Chen Wu ◽  
Michael Lichten
Keyword(s):  
Meiotic Recombination ◽  
Double Strand Breaks ◽  
Target Sequence ◽  
Dna Double Strand Breaks ◽  
Cold Region ◽  
Independent Manner ◽  
Strand Breaks ◽  
Intergenic Regions ◽  
Chromosome Iii ◽  
The Right

ABSTRACT In Saccharomyces cerevisiae, meiotic recombination is initiated by DNA double-strand breaks (DSBs). DSBs usually occur in intergenic regions that display nuclease hypersensitivity in digests of chromatin. DSBs are distributed nonuniformly across chromosomes; on chromosome III, DSBs are concentrated in two “hot” regions, one in each chromosome arm. DSBs occur rarely in regions within about 40 kb of each telomere and in an 80-kb region in the center of the chromosome, just to the right of the centromere. We used recombination reporter inserts containing arg4 mutant alleles to show that the “cold” properties of the central DSB-deficient region are imposed on DNA inserted in the region. Cold region inserts display DSB and recombination frequencies that are substantially less than those seen with similar inserts in flanking hot regions. This occurs without apparent change in chromatin structure, as the same pattern and level of DNase I hypersensitivity is seen in chromatin of hot and cold region inserts. These data are consistent with the suggestion that features of higher-order chromosome structure or chromosome dynamics act in a target sequence-independent manner to control where recombination events initiate during meiosis.

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 Mating type–dependent constraints on the mobility of the left arm of yeast chromosome III

2004 ◽  
Vol 164 (3) ◽  
pp. 361-371 ◽  
Author(s):  
Debra A. Bressan ◽  
Julio Vazquez ◽  
James E. Haber
Keyword(s):  
Mating Type ◽  
Three Dimensional ◽  
Cell Mobility ◽  
Yeast Chromosome ◽  
Mating Type Gene ◽  
Type Gene ◽  
Three Dimensional Configuration ◽  
Chromosome Iii ◽  
The Right ◽  
Donor Preference

Mating-type gene (MAT) switching in budding yeast exhibits donor preference. MATa preferentially recombines with HML near the left telomere of chromosome III, whereas MATα prefers HMR near the right telomere. Donor preference is controlled by the recombination enhancer (RE) located proximal to HML. To test if HML is constrained in pairing with MATα, we examined live-cell mobility of LacI-GFP–bound lactose operator (lacO) arrays inserted at different chromosomal sites. Without induction of recombination, lacO sequences adjacent to HML are strongly constrained in both MATα and RE-deleted MATa strains, compared with MATa. In contrast, chromosome movement at HMR or near a telomere of chromosome V is mating-type independent. HML is more constrained in MATa Δre and less constrained in MATa RE+ compared with other sites. Although HML and MATa are not prealigned before inducing recombination, the three-dimensional configuration of MAT, HML, and HMR is mating-type dependent. These data suggest there is constitutive tethering of HML, which is relieved in MATa cells through the action of RE.

scholarly journals Xist Yeast Artificial Chromosome Transgenes Function as X-Inactivation Centers Only in Multicopy Arrays and Not as Single Copies

1999 ◽  
Vol 19 (4) ◽  
pp. 3156-3166 ◽  
Author(s):  
Edith Heard ◽  
Fabien Mongelard ◽  
Danielle Arnaud ◽  
Philip Avner
Keyword(s):  
X Chromosome ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Diploid Cell ◽  
Single Copy ◽  
X Inactivation ◽  
Artificial Chromosomes ◽  
Autonomous Function ◽  
Inactivation Center ◽  
Yeast Artificial Chromosomes

ABSTRACT X-chromosome inactivation in female mammals is controlled by the X-inactivation center (Xic). This locus is required for inactivation incis and is thought to be involved in the counting process which ensures that only a single X chromosome remains active per diploid cell. The Xist gene maps to the Xic region and has been shown to be essential for inactivation in cis. Transgenesis represents a stringent test for defining the minimal region that can carry out the functions attributed to the Xic. Although YAC and cosmid Xist-containing transgenes have previously been reported to be capable of cis inactivation and counting, the transgenes were all present as multicopy arrays and it was unclear to what extent individual copies are functional. Using two different yeast artificial chromosomes (YACs), we have found that single-copy transgenes, unlike multicopy arrays, can induce neither inactivation in cis nor counting. These results demonstrate that despite their large size and the presence of Xist, the YACs that we have tested lack sequences critical for autonomous function with respect to X inactivation.

scholarly journals Meiotic recombination between yeast artificial chromosomes yields a single clone containing the entire BCL2 protooncogene.

1990 ◽  
Vol 87 (24) ◽  
pp. 9913-9917 ◽  
Author(s):  
G. A. Silverman ◽  
E. D. Green ◽  
R. L. Young ◽  
J. I. Jockel ◽  
P. H. Domer ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Artificial Chromosomes ◽  
Single Clone ◽  
Yeast Artificial Chromosomes

scholarly journals Polymorphisms on the right arm of yeast chromosome III associated with Ty transposition and recombination events

1987 ◽  
Vol 15 (21) ◽  
pp. 8963-8982 ◽  
Author(s):  
John R. Warmington ◽  
Rebecca P. Green ◽  
Carol S. Newlon ◽  
Stephen G. Oliver
Keyword(s):  
Yeast Chromosome ◽  
Chromosome Iii ◽  
The Right
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