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Open AccessJournal ArticleDOI

Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae.

Brian Sauer
- 01 Jun 1987 - 
- Vol. 7, Iss: 6, pp 2087-2096
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TLDR
It is demonstrated that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recomb inase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure.
Abstract
The procaryotic cre-lox site-specific recombination system of coliphage P1 was shown to function in an efficient manner in a eucaryote, the yeast Saccharomyces cerevisiae. The cre gene, which codes for a site-specific recombinase, was placed under control of the yeast GALI promoter. lox sites flanking the LEU2 gene were integrated into two different chromosomes in both orientations. Excisive recombination at the lox sites (as measured by loss of the LEU2 gene) was promoted efficiently and accurately by the Cre protein and was dependent upon induction by galactose. These results demonstrate that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recombinase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure.

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Citations
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Journal ArticleDOI

A New Efficient Gene Disruption Cassette for Repeated Use in Budding Yeast

TL;DR: A loxP-kanMX-loxP gene disruption cassette is developed, which combines the advantages of the heterologous kanr marker with those from the Cre- lox P recombination system, and will be of great advantage for the functional analysis of gene families.
Journal ArticleDOI

Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting.

TL;DR: This work has employed a method based on the Cre-loxP recombination system of bacteriophage P1 to generate a mouse strain in which the JH segments and the intron enhancer in the IgH locus are deleted, and shows that, on the mutant chromosome, switch recombination at the mu gene switch region is strongly suppressed, whereas the switch region of the gamma 1 gene is efficiently rearranged.
Journal ArticleDOI

The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome.

TL;DR: The site-specific recombination system of the yeast 2 micron plasmid, the FLP recombinase and its recombination targets (FRTs), into the genome of Drosophila, producing white-eyed and dark-red-eyed progeny.
Journal ArticleDOI

Construction of adenovirus vectors through Cre-lox recombination.

TL;DR: First, a simple method for constructing E1-substituted adenovirus vectors carrying recombinant genes in place of all of the viral genes is demonstrated, so-called gutless adenOVirus vectors.
References
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Journal ArticleDOI

Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal ArticleDOI

A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Book

Molecular Cloning: A Laboratory Manual

TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal Article

Cleavage of structural proteins during the assemble of the head of bacterio-phage T4

U. K. Laemmli
- 01 Jan 1970 - 
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Journal ArticleDOI

DNA sequencing with chain-terminating inhibitors

TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
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