Nucleotide sequence encoding the enzyme I-SecI and the uses thereof

• US 6,833,252 B1
• Filed: 01/27/2000
• Issued: 12/21/2004
• Est. Priority Date: 05/05/1992
• Status: Expired due to Fees

• Current Assignee: Institut Pasteur, University Pierre et Marie Curie
• Original Assignee: Institut Pasteur, University Pierre et Marie Curie
• Inventors: Dujon, Bernard, Nicolas, Jean-Francois, Choulika, Andre, Perrin, Arnaud
• Primary Examiner(s): Fredman, Jeffrey
• Assistant Examiner(s): Kaushal, Sumesh
• Application Number:
  US09/492,697
• Time in Patent Office: 1,790 Days
• Field of Search: 435/69.1, 435/320.1, 435/325, 435/455, 435/354, 536/23.1, 536/23.74, 536/24.1
• US Class Current: 435/69.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: C12N 15/66 General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Q 1/683 involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: Y10S 530/824 Yeasts

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.

Claims

1. A recombinant mammalian chromosome comprising an exogenous Group I intron encoded endonuclease site,

wherein the endonuclease site is within an integrated nucleic acid sequence from a vector,

wherein the site is selected from the group consisting of an I-SceIV site, an I-CsmI site, I-PanI site, I-SceII site, an I-CeuI site, an I-PpoI site, an I-SceIII site, an I-CreI site, an I-TevI site, an I-TevII site, an I-TevIII site, and an I-SceI site.

2. The recombinant mammalian chromosome of claim 1, wherein the site is an I-SceIV site.

3. The recombinant mammalian chromosome of claim 1, wherein the site is an I-CsmI site.

4. The recombinant mammalian chromosome of claim 1, wherein the site is an I-PanI site.

5. The recombinant mammalian chromosome of claim 1, wherein the site is an I-SceII site.

6. The recombinant mammalian chromosome of claim 1, wherein the site is an I-CeuI site.

7. The recombinant mammalian chromosome of claim 1, wherein the site is an I-PpoI site.

8. The recombinant mammalian chromosome of claim 1, wherein the site is an I-SceIII site.

9. The recombinant mammalian chromosome of claim 1, wherein the site is an I-CreI site.

10. The recombinant mammalian chromosome of claim 1, wherein the site is an I-TevI site.

11. The recombinant mammalian chromosome of claim 1, wherein the site is an I-TevII site.

12. The recombinant mammalian chromosome of claim 1, wherein the site is an I-TevIII site.

13. The recombinant mammalian chromosome of claim 1, wherein the site is an I-SceI site.

14. The recombinant mammalian chromosome of any of claims 1-13, wherein the chromosome is a murine chromosome.

15. The recombinant mammalian chromosome of any of claims 1-13, wherein the chromosome is a mouse chromosome.

16. A recombinant cell comprising the recombinant chromosome of any of claims 1-13.

17. A recombinant cell comprising the recombinant chromosome of claim 14.

18. A recombinant cell comprising the recombinant chromosome of claim 15.

19. The recombinant cell of claim 18, wherein the recombinant cell is a mouse stem cell.