NESTED CHROMOSOMAL FRAGMENTATION IN YEAST USING THE MEGANUCLEASE I-SCE-I - A NEW METHOD FOR PHYSICAL MAPPING OF EUKARYOTIC GENOMES

被引：69

作者：

THIERRY, A

DUJON, B

机构：

[1] Unité de Génétique Moléculaire des Levures, Département de Biologie Moléculaire (URA 1149 du CNRS), Institut Pasteur, F-75724 Paris-Cedex 15

来源：

NUCLEIC ACIDS RESEARCH | 1992年 / 20卷 / 21期

关键词：

D O I：

10.1093/nar/20.21.5625

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 have developed a new method for the physical mapping of genomes and the rapid sorting of genomic libraries which is based on chromosome fragmentation by the meganuclease I-Sce I, the first available member of a new class of endonucleases with very long recognition sequences. I-Sce I allows complete cleavage at a single artificially inserted site in an entire genome. Sites can be inserted by homologous recombination using specific cassettes containing selectable markers or, at random, using transposons. This method has been applied to the physical mapping of chromosome XI (620 kb) of Saccharomyces cerevisi and to the sorting of a cosmid library. Our strategy has potential applications to various genome mapping projects. A set of transgenic yeast strains carrying the I-Sce I sites at various locations along a chromosome defines physical intervals against which new genes, DNA fragments or clones can be mapped directly by simple hybridizations.

引用

页码：5625 / 5631

页数：7

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