Isolation of an 800 kb contiguous DNA fragment encompassing a 3.5-cM region of chromosome 1 in Arabidopsis using YAC clones

Genome ◽  
1995 ◽  
Vol 38 (4) ◽  
pp. 817-823 ◽  
Author(s):  
Usha Vijayraghavan ◽  
Imran Siddiqi ◽  
Elliot Meyerowitz
Keyword(s):  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Chromosome 1 ◽  
Rflp Markers ◽  
Genomic Libraries ◽  
Physical Maps ◽  
Starting Point ◽  
Meristem Identity ◽  
Dna Fragment

The apetala1 mutation of Arabidopsis affects floral meristem identity and the development of sepal and petal primordia of the flower. We mapped the available RFLP markers on chromosome 1 that are in the general vicinity of apetala1 on a fine structure map and then chose the closest RFLP as a starting point for contiguous DNA (contig) generation. We report here a contig of about 800 kilobases (kb) that spans a 3.5 cM region of chromosome 1. We used genomic libraries of Arabidopsis prepared in yeast artificial chromosome (YAC) vectors and the detailed characterization of 19 YACs is reported. RFLPs displayed by the end fragments from the walk were mapped to align and correlate the genetic and physical maps for this region of chromosome 1. In this segment of the genome, 1 cM corresponds to a little over 200 kb of physical distance.Key words: Arabidopsis, apetala1, chromosome walking.

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.

Evolving strategies for making physical maps of mammalian chromosomes

Genome ◽  
1989 ◽  
Vol 31 (2) ◽  
pp. 1055-1058 ◽  
Author(s):  
Cassandra L. Smith ◽  
Charles R. Cantor
Keyword(s):  
Yeast Artificial Chromosome ◽  
Polymerase Chain Reaction Amplification ◽  
Artificial Chromosome ◽  
Dna Polymorphisms ◽  
Cleavage Sites ◽  
Restriction Maps ◽  
Physical Maps ◽  
Mammalian Genomes ◽  
Reaction Amplification ◽  
Polymerase Chain

Two types of physical maps are described: restriction maps made by top down approaches using enzymes that cut the genome infrequently, and complete libraries, made by bottom up approaches using fingerprinting of randomly selected cloned DNA. Construction of such maps for mammalian chromosomes is complicated by the mosaic nature of mammalian genomes, and extensive polymorphisms at the cleavage sites of most enzymes that yield large DNA fragments. However, it appears that both of these potential difficulties can be turned into advantages by new mapping strategies. When combined with yeast artificial chromosome cloning and polymerase chain reaction amplification methods, these approaches should soon yield complete maps of many human chromosomes.Key words: fingerprinting, restriction maps, DNA polymorphisms, human genome.

Characterization of human genomic yeast artificial chromosome inserts containing hexokinase 1 coding information on chromosome 10

1992 ◽  
Vol 47 (3) ◽  
pp. 265-269
Author(s):  
Bruce D. Gelb ◽  
Kim C. Worley ◽  
Lisa D. Griffin ◽  
Volker Admas ◽  
A.Craig Chinault ◽  
...  
Keyword(s):  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Chromosome 10 ◽  
Human Genomic

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.

Transgenic Analysis of a 100-kb Human β-Globin Cluster–Containing DNA Fragment Propagated as a Bacterial Artificial Chromosome

Blood ◽  
1999 ◽  
Vol 94 (9) ◽  
pp. 3178-3184 ◽  
Author(s):  
Richard M. Kaufman ◽  
Christine T.N. Pham ◽  
Timothy J. Ley
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Yeast Artificial Chromosome ◽  
Globin Gene ◽  
Artificial Chromosome ◽  
Pcr Analysis ◽  
Expression Levels ◽  
Dna Fragment ◽  
Polymerase Chain ◽  
Gene Switching ◽  
Intermediate Expression

To date, the normal transcriptional regulation of the human β-globin gene cluster has been recapitulated most accurately in transgenic mice that carry large yeast artificial chromosome (YAC) or ligated cosmid constructs. However, these large transgenes still exhibit variegated expression levels, perhaps because they tend to rearrange upon integration, or because the cloning vectors remain attached to the globin inserts. To try to circumvent these potential problems, we investigated the transgenic properties of a 100-kb DNA fragment containing the entire human β-globin cluster propagated in a bacterial artificial chromosome (BAC). We created 9 independent mouse lines, each carrying 1 to 6 copies of the human β-globin cluster without the attached BAC vector. Five of the lines carry unrearranged copies of the cluster. Reverse-transcriptase polymerase chain reaction (RT-PCR) analysis of adult F1 mice showed that 2 lines express human β globin at levels approximately equivalent to the endogenous mouse β-major genes. One line expresses no human β globin, while the remaining 6 lines show intermediate expression levels. Complete γ→β-globin gene switching occurs, but is slightly delayed with respect to the endogenous mouse embryonic→adult switch. Since these data are similar to what has been obtained using globin YACs or ligated cosmids, we conclude that (1) globin transgenes propagated in BACs are no less likely to rearrange than their cosmid or YAC counterparts, and (2) the retention of YAC vector sequences in a transgene probably has no significant impact on globin expression when using constructs of this size.

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