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PMID: 2102834 Published · ppublish English Journal Article

Isolation of single-copy-sequence clones from a yeast artificial chromosome library of randomly-sheared Arabidopsis thaliana DNA.

Plant molecular biology ·Vol. 14 ·No. 4 ·1990-04-00 ·Pages 561-8

Ward ER, Jen GC

Abstract

We describe the construction of a yeast artificial chromosome (YAC) library from the Arabidopsis thaliana genome. Randomly sheared high molecular weight source DNA was extracted from frozen, ground leaf tissue and blunt-end-ligated to the vector pYAC3. By size-fractionating the ligation products, we achieved an average clone size of 150 kb. Approximately 6% of the YACs contained inserts from the chloroplast genome. We screened clones equivalent to greater than four A. thaliana haploid nuclear genomes and isolated YACs homologous to five single-copy-sequence probes. The library should be useful for chromosome walking and genome mapping experiments. In addition, the approach used for its construction should be applicable to other higher plant species.

MeSH Terms
Chromosomes Cloning, Molecular DNA/genetics,isolation & purification Gene Library Genetic Vectors Plants/genetics Saccharomyces cerevisiae/genetics
Chemicals
DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ward E R
Agricultural Biotechnology Research Unit, CIBA-GEIGY Corporation, Research Triangle Park, NC 27709-2257.
Jen G C
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
DOI
10.1007/BF00027501
Published
1990-04-00
Pages
561-8
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00027501 View journal details
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