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US 5,474,896 AGranted
Nucleotide sequence encoding the enzyme I-SceI and the uses thereof
AssigneesInstitut Pasteur, Universite Paris-VI
InventorsBernard Dujon, Andre Choulika, Laurence Colleaux, Cecile Fairhead, Arnaud Perrin, Anne Plessis, Agnes Thierry
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Abstract
An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.
Claims2
A method of genetically mapping a yeast genome that does not contain a natural restriction site for I-SceI, comprising the steps of:
- (a) artificially inserting one or more I-SceI sites at various positions in the genome;
- (b) completely cleaving said genome at the inserted I-SceI sites, with the restriction enzyme I-SceI, to produce nested chromosomal fragments;
- (c) purifying said fragments of step (b) by pulsed field gel electrophoresis;
- (d) transferring the fragments to a solid membrane;
- (e) hybridizing the fragments bound to said membrane to a labelled probe derived from a cosmid clone, pUKG040.
- (f) detecting the hybridization banding patterns;
- and
- (g) mapping said yeast genome based on the hybridization banding patterns observed in step (f).
5 more elements
A method of genetically mapping a yeast genome that does not contain a natural restriction site for I-SceI, comprising the steps of:
- (a) artificially inserting one or more I-SceI sites at various positions in the genome;
- (b) completely cleaving said genome at the inserted I-SceI sites, with the restriction enzyme I-SceI, to produce nested chromosomal fragments;
- (c) purifying said fragments of step (b) by pulsed field gel electrophoresis;
- (d) transferring the fragments to a solid membrane;
- (e) hybridizing the fragments bound to said membrane to a labelled probe derived from a cosmid clone, pUKG066.
- (f) detecting the hybridization banding patterns;
- and
- (g) mapping said yeast genome based on the hybridization banding patterns observed in step (f).
5 more elements
Dependencies are inferred from the claim text.