Construction of an Overlapping YAC Library of the Arabidopsis thaliana Genome

As part of an international effort to construct an overlapping yeast artificial chromosome (YAC) library of the Arabidopsis thaliana genome, we have been identifying and linking YAC clones that cover the top halves of chromosomes 4 and 5. All the available RFLP markers mapping to these regions were hybridised to filters carrying different YAC libraries. The YAC colonies were arrayed such that an entire library was contained on one small filter, greatly simplifying the screening procedure. The collaborative effort has currently resulted in 30% of the genome being represented in mapped YAC clones. Probes derived from the ends of the YAC inserts, using either inverse polymerase chain reaction (PCR) or left-end rescue, are now being used to walk out to link the YAC clones hybridising to adjacent RFLP markers.

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Analysis of Yeast Artificial Chromosome DNA by Restriction Digestion, Southern Blotting Nucleic Acid Hybridization, and Polymerase Chain Reaction

YAC Protocols ◽

10.1385/1-59745-158-4:33 ◽

2006 ◽

pp. 33-40

Author(s):

Sylvia Vasiliou ◽

John Peter Quinn

Keyword(s):

Polymerase Chain Reaction ◽

Nucleic Acid ◽

Southern Blotting ◽

Yeast Artificial Chromosome ◽

Artificial Chromosome ◽

Nucleic Acid Hybridization ◽

Restriction Digestion ◽

Chain Reaction ◽

Polymerase Chain

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Alu-primed polymerase chain reaction for regional assignment of 110 yeast artificial chromosome clones from the human X chromosome: identification of clones associated with a disease locus.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.88.14.6157 ◽

1991 ◽

Vol 88 (14) ◽

pp. 6157-6161 ◽

Cited By ~ 30

Author(s):

D. L. Nelson ◽

A. Ballabio ◽

M. F. Victoria ◽

M. Pieretti ◽

R. D. Bies ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

X Chromosome ◽

Yeast Artificial Chromosome ◽

Artificial Chromosome ◽

Disease Locus ◽

Chromosome Identification ◽

Chain Reaction ◽

Polymerase Chain ◽

Regional Assignment

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Simple and robust screening of pooled yeast artificial chromosome libraries by the restriction enzyme digestion of polymerase chain reaction products

Genetic Analysis Biomolecular Engineering ◽

10.1016/1050-3862(94)90052-3 ◽

1994 ◽

Vol 11 (3) ◽

pp. 63-68 ◽

Cited By ~ 1

Author(s):

Xueqian Wang ◽

Jing Qian ◽

Minoru S.H. Ko

Keyword(s):

Polymerase Chain Reaction ◽

Restriction Enzyme ◽

Yeast Artificial Chromosome ◽

Artificial Chromosome ◽

Enzyme Digestion ◽

Restriction Enzyme Digestion ◽

Reaction Products ◽

Chain Reaction ◽

Polymerase Chain

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

Genome ◽

10.1139/g96-136 ◽

1996 ◽

Vol 39 (6) ◽

pp. 1086-1092 ◽

Cited By ~ 5

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.

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

Functional Plant Biology ◽

10.1071/pp9960453 ◽

1996 ◽

Vol 23 (4) ◽

pp. 453 ◽

Cited By ~ 3

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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