ISOLATION OF SINGLE-COPY-SEQUENCE CLONES FROM A YEAST ARTIFICIAL CHROMOSOME LIBRARY OF RANDOMLY-SHEARED ARABIDOPSIS-THALIANA DNA

被引：121

作者：

WARD, ER ^{[1
]}

JEN, GC ^{[1
]}

机构：

[1] CIBA GEIGY CORP,AGR BIOTECHNOL RES UNIT,POB 12257,RES TRIANGLE PK,NC 27709

来源：

PLANT MOLECULAR BIOLOGY | 1990年 / 14卷 / 04期

关键词：

chromosome walking; genome mapping; plant genetics; YAC;

D O I：

10.1007/BF00027501

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 chromosome walking and genome mapping experiments. In addition, the approach used for its construction should be applicable to other higher plant species. © 1990 Kluwer Academic Publishers.

引用

页码：561 / 568

页数：8

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