scholarly journals REMI-RFLP mapping in the Dictyostelium genome.

Genetics ◽  
1994 ◽  
Vol 138 (3) ◽  
pp. 665-674 ◽  
Author(s):  
A Kuspa ◽  
W F Loomis
Keyword(s):  
Yeast Artificial Chromosome ◽  
Fragment Length Polymorphism ◽  
Physical Map ◽  
Artificial Chromosome ◽  
Genome Alignment ◽  
Hybridization Probes ◽  
Restriction Sites ◽  
Mapping Techniques ◽  
Yac Contig ◽  
Restriction Fragment Length

Abstract A set of 147 Dictyostelium discoideum strains was constructed by random integration of a vector containing rare restriction sites. The strains were generated by transformation using restriction enzyme-mediated integration (REMI) which results in the integration of linear DNA fragments into randomly distributed genomic restriction sites. Restriction fragment length polymorphism (RFLP) was generated in a single genomic site in each strain. These REMI-RFLP strains were used to confirm gene linkages previously supported by two other physical mapping techniques: yeast artificial chromosome (YAC) contig construction, and megabase-scale restriction mapping. New linkages were uncovered when two or more hybridization probes identified the same RFLP fragments. Probes for 100 genes have marked 53% of the RFLPs, representing greater than 22 Mb of the 40 Mb Dictyostelium genome. Alignment of these and other large fragments along each chromosome should lead to a complete physical map of the Dictyostelium genome.

scholarly journals Structure of the Chromosome VII Centromere Region in Neurospora crassa: Degenerate Transposons and Simple Repeats

1998 ◽  
Vol 18 (9) ◽  
pp. 5465-5477 ◽  
Author(s):  
Edward B. Cambareri ◽  
Rafael Aisner ◽  
John Carbon
Keyword(s):  
Neurospora Crassa ◽  
Yeast Artificial Chromosome ◽  
Fragment Length Polymorphism ◽  
Artificial Chromosome ◽  
Polymorphism Analysis ◽  
Dna Repeats ◽  
Centromere Region ◽  
Simple Repeats ◽  
Restriction Fragment Length ◽  
Repeat Induced Point Mutation

ABSTRACT DNA from the centromere region of linkage group (LG) VII ofNeurospora crassa was cloned previously from a yeast artificial chromosome library and was found to be atypical ofNeurospora DNA in both composition (AT rich) and complexity (repetitive). We have determined the DNA sequence of a small portion (∼16.1 kb) of this region and have identified a cluster of three new retrotransposon-like elements as well as degenerate fragments from the 3′ end of Tad, a previously identified LINE-like retrotransposon. This region contains a novel full-length but nonmobilecopia-like element, designated Tcen, that is only associated with centromere regions. Adjacent DNA contains portions of a gypsy-like element designated Tgl1. A third new element, Tgl2, shows similarity to theTy3 transposon of Saccharomyces cerevisiae. All three of these elements appear to be degenerate, containing predominantly transition mutations suggestive of the repeat-induced point mutation (RIP) process. Three new simple DNA repeats have also been identified in the LG VII centromere region. While Tcenelements map exclusively to centromere regions by restriction fragment length polymorphism analysis, the defective Tad elements appear to occur most frequently within centromeres but are also found at other loci including telomeres. The characteristics and arrangement of these elements are similar to those seen in theDrosophila centromere, but the relative abundance of each class of repeats, as well as the sequence degeneracy of the transposon-like elements, is unique to Neurospora. These results suggest that the Neurospora centromere is heterochromatic and regional in character, more similar to centromeres of Drosophila than to those of most single-cell yeasts.

A physical map with yeast artificial chromosome (YAC) clones covering 63% of the 12 rice chromosomes

Genome ◽  
2001 ◽  
Vol 44 (1) ◽  
pp. 32-37 ◽  
Author(s):  
Shoko Saji ◽  
Yosuke Umehara ◽  
Baltazar A Antonio ◽  
Hiroko Yamane ◽  
Hiroshi Tanoue ◽  
...  
Keyword(s):  
Genetic Map ◽  
Dna Markers ◽  
Yeast Artificial Chromosome ◽  
Physical Map ◽  
Genome Structure ◽  
Rice Genome ◽  
Artificial Chromosome ◽  
Physical Length ◽  
Rice Chromosomes ◽  
Yac Contig

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.

A 2.8 megabase YAC contig spanning D8S339, which is tightly linked to the Werner syndrome locus

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 77-83
Author(s):  
R. Bruskiewich ◽  
M. Schertzer ◽  
S. Wood
Keyword(s):  
Human Chromosome ◽  
Genetic Marker ◽  
Yeast Artificial Chromosome ◽  
Werner Syndrome ◽  
Physical Map ◽  
Artificial Chromosome ◽  
Sequence Tagged Sites ◽  
Yeast Artificial Chromosome Contig ◽  
Yac Contig ◽  
Chromosome 8P

A number of gene loci, including the locus for Werner syndrome (WRN), map to proximal human chromosome 8p near the genetic marker D8S339. In this report, we present a long range physical map of an approximately 2.8 megabase yeast artificial chromosome contig centred on D8S339. In this map, we localize the WRN-linked polymorphic sequence-tagged sites (STS) D8S339 and D8S1055, as well as a novel polymorphic STS, D8S2297. We also refine the positions of three known gene loci, GTF2E2, GSR, and PPP2CB, relative to the location of WRN within the map.Key words: WRN, GTF2E2, GSR, PPP2CB, physical map, human chromosome 8p.

Genetic and contig map of a 2200-kb region encompassing 5.5 cM on chromosome 1 of Arabidopsis thaliana

Genome ◽  
1996 ◽  
Vol 39 (6) ◽  
pp. 1086-1092 ◽  
Author(s):  
Christian S. Hardtke ◽  
Thomas Berleth
Keyword(s):  
Arabidopsis Thaliana ◽  
Positional Cloning ◽  
Yeast Artificial Chromosome ◽  
Physical Map ◽  
Primary Root ◽  
Artificial Chromosome ◽  
Chromosome 1 ◽  
Rflp Markers ◽  
Caps Markers ◽  
Yac Contig

In the course of the isolation of the MONOPTEROS (MP) gene, required for primary root formation in Arabidopsis thaliana, a yeast artificial chromosome (YAC) contig encompassing approximately 2200 kilobases corresponding to 5.5 cM on the top arm of chromosome 1 was established. Forty-six YAC clones were characterized and 12 new restriction fragment length polymorphism (RFLP) markers are presented. Three new codominant amplified polymorphic sequence (CAPS) markers were generated that enabled high resolution genetic mapping and correlation of physical and genetic distances along the contig. The map contributes to the completion of a physical map of the Arabidopsis genome and should facilitate positional cloning of other genes in the region as well as studies on genome organization. We also present another set of 11 physically linked probes, as well as mapping data for additional RFLP markers within a broader interval of 10.4 cM. Key words : Arabidopsis, CAPS markers, MONOPTEROS gene, physical map, RFLP markers, YAC contig.

scholarly journals The CMT2D Locus: Refined Genetic Position and Construction of a Bacterial Clone-Based Physical Map

1999 ◽  
Vol 9 (6) ◽  
pp. 568-574 ◽  
Author(s):  
Rachel E. Ellsworth ◽  
Victor Ionasescu ◽  
Charles Searby ◽  
Val C. Sheffield ◽  
Valerie V. Braden ◽  
...  
Keyword(s):  
Linkage Analysis ◽  
Yeast Artificial Chromosome ◽  
Sequence Data ◽  
Physical Map ◽  
Muscular Atrophy ◽  
Large Family ◽  
Artificial Chromosome ◽  
Single Chromosome ◽  
Mixed Features ◽  
Bacterial Clone

Charcot-Marie-Tooth (CMT) disease is a progressive neuropathy of the peripheral nervous system, typically characterized by muscle weakness of the distal limbs. CMT is noted for its genetic heterogeneity, with four distinct loci already identified for the axonal form of the disease (CMT2). In 1996, linkage analysis of a single large family revealed the presence of a CMT2 locus on chromosome 7p14 (designatedCMT2D). Additional families have been linked subsequently to the same genomic region, including one with distal spinal muscular atrophy (dSMA) and one with mixed features of dSMA and CMT2; symptoms in both of these latter families closely resemble those seen in the original CMT2D family. There is thus a distinct possibility that CMT2 and dSMA encountered in these families reflect allelic heterogeneity at a single chromosome 7 locus. In the study reported here, we have performed more detailed linkage analysis of the original CMT2D family based on new knowledge of the physical locations of various genetic markers. The region containing the CMT2D gene, as defined by the original family, overlaps with those defined by at least two other families with CMT2 and/or dSMA symptoms. Both yeast artificial chromosome (YAC) and bacterial clone-based [bacterial artificial chromosome (BAC) and P1-derived artificial chromosome (PAC)] contig maps spanning ∼3.4 Mb have been assembled across the combinedCMT2D critical region, with the latter providing suitable clones for systematic sequencing of the interval. Preliminary analyses have already revealed at least 28 candidate genes and expressed-sequence tags (ESTs). The mapping information reported here in conjunction with the evolving sequence data should expedite the identification of the CMT2D/dSMA gene or genes.

scholarly journals Dosage and Imprinting Effects in Abnormalities of Human Chromosome 15

1996 ◽  
Vol 45 (1-2) ◽  
pp. 83-83 ◽  
Author(s):  
D.H. Ledbetter ◽  
S.L. Christian ◽  
T. Kubota ◽  
A. Mutirangura ◽  
J.S. Sutcliffe ◽  
...  
Keyword(s):  
Yeast Artificial Chromosome ◽  
Cpg Island ◽  
Uniparental Disomy ◽  
Artificial Chromosome ◽  
Chromosome 15 ◽  
Diagnostic Strategy ◽  
Str Analysis ◽  
Paternal Grandmother ◽  
Cosmid Contigs ◽  
Yac Contig

Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are distinct mental retardation disorders caused by paternal deficiency (PWS) or maternal deficiency (AS) of gene(s) in 15qll.2-ql3. We have constructed a 3.5 Mb yeast artificial chromosome (YAC) contig of the PWS/AS region and cosmid contigs of selected YACs at D15S13, SNRPN, S10, and S113. Cosmid clones have been used for fluorescence in situ hybridization (FISH) detection of deletions in PWS and AS patients. In addition, a total of 28 short tandem repeat polymorphisms (STRs) have been mapped to specific YACs in the contig, providing a highly informative set of markers for detection of deletion or uniparental disomy (UPD) in PWS and AS patients. Use of the 3 most informative markers in this region (S542, S128, and ASSCA-1) plus 3 markers distal on 15q (S123, S125, and S131) provide an efficient diagnostic strategy for UPD15.A combination of FISH and STR analysis has identified small deletions in one sporadic and one familial case of PWS (family O). Both deletions involve all or part of the SNRPN gene but do not extend telomeric to PAR-5 or PAR-1, two novel transcripts expressed exclusively from the paternal chromosome. However, expression of SNRPN, PAR-5, and PAR-1 is lost in both cases, implying the presence of an imprinting control region near SNRPN. The smallest deletion in family O is estimated at approximately 30-40 kb in size and involves a newly identified CpG island at the 5′ end of SNRPN which is methylated on the maternal chromosome. This small deletion in two PWS affected siblings was present in the father and the paternal grandmother, both of whom were phenotypically normal.

A 6-Mb Yeast Artificial Chromosome Contig and Long-Range Physical Map Encompassing the Region on Chromosome 12q15 Frequently Rearranged in a Variety of Benign Solid Tumors

Genomics ◽  
1995 ◽  
Vol 29 (3) ◽  
pp. 665-678 ◽  
Author(s):  
ERIC F.P.M. SCHOENMAKERS ◽  
JAN M.W. GEURTS ◽  
PATRICK F.J. KOOLS ◽  
RAF MOLS ◽  
CHRISTEL HUYSMANS ◽  
...  
Keyword(s):  
Solid Tumors ◽  
Long Range ◽  
Yeast Artificial Chromosome ◽  
Physical Map ◽  
Artificial Chromosome ◽  
Yeast Artificial Chromosome Contig

scholarly journals MOLECULAR IDENTIFICATION AND CLONAL RELATION OF ATYPICAL ISOLATE Aeromonas salmonicida USING RESTRICTION FRAGMENT LENGTH POLYMORPHISM (RFLP)

2011 ◽  
Vol 6 (2) ◽  
pp. 183
Author(s):  
Riza Priyatna ◽  
Indaryulianto Indaryulianto ◽  
Kurniasih Kurniasih
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Aeromonas Salmonicida ◽  
Freshwater Aquaculture ◽  
Restriction Sites ◽  
Restriction Fragment Length

Aeromonas salmonicida is responsible in many cases of furunculosis outbreaks resulting in economic loss of freshwater aquaculture. Four isolates of A. salmonicida have been isolated from fish collected in four different regions in Indonesia and its clonal relation has yet to be determined. In the research, A. salmonicida isolates and ATCC atypical isolate as the control had been checked for their clonal relation using Restriction Fragment Length Polymorphism (RFLP) method in which restriction enzyme of AluI, HaeIII, MboI, and EheI were used. PCR test results using the primers 16S rDNA amplicon gave a positive response to the 1300 bp band. The result of RFLP analysis showed that A. salmonicida atypical isolates from Indonesia represent subspecies smithia except isolates from C. macropomum in Yogyakarta in MS and 16S rDNA regions. Isolates from Jambi, Pontianak and Semarang showed a slight variation on enzyme restriction sites. Isolates number 2, 3, 4 and 5 had the same restriction sites using AluI enzyme with MS primer. The restriction enzymes that could give the best result for RFLP method of A. salmonicida were HaeIII, MboI and EheI.

Construction of a YAC Contig of 2 Megabases Around the MS1 Gene in Arabidopsis thaliana

1996 ◽  
Vol 23 (4) ◽  
pp. 453 ◽  
Author(s):  
RM Chapple ◽  
AM Chaudhury ◽  
KC Blomer ◽  
LB Farrell ◽  
ES Dennis
Keyword(s):  
Arabidopsis Thaliana ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Rflp Markers ◽  
Chromosome Walking ◽  
Chromosome 5 ◽  
Plant Populations ◽  
Recombinant Plant ◽  
Yac Contig ◽  
The Relationship

The ms1 mutation of Arabidopsis thaliana causes male sterility by preventing the development of normal microspores in the developing anther. The gene is located on a region of chromosome 5 containing the RFLP markers g4111, g4560 and g21503. Using yeast artificial chromosome (YAC) libraries, we have constructed a contig of 38 YACs spanning approximately 2.1 megabases (approximately 2% of the genome) around MS1 and redefined the order of these RFLP markers. Chimeric YACs and repetitive DNA caused problems in chromosome 'walking'. A method for cloning YAC right ends by plasmid rescue was applied to arabidopsis. One YAC end contained a portion of the A. thaliana sucrose synthase gene ASUSI, hence locating this gene on chromosome 5 near MS1. Using recombinant plant populations containing ms1 and flanking markers, MS1 was localised to a 200 kb region within the YAC contig. In this contig the relationship between physical and genetic distance varied from less than 100 kb to 720 kb per centimorgan.

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