Use of beta recombinase in eukaryotic cells, especially for transgenic work

Abstract

The invention relates to the use of prokaryotic beta recombinase in eukaryotic cells, especially for transgenic work in eukaryotic cells. It also relates to the use of prokaryotic beta recombinase for site-specific intramolecular recombination between two six sites in eukaryotic cells. The use of the gene coding for beta recombinase for promoting the deletion of DNA sequences located between directly oriented six sites in mammalian cells and for catalysing site-specific resolution of DNA sequences in an extrachromosomal target introduced into an eukaryotic cell are also disclosed.

1 . Use of prokaryotic beta recombinase and its specific target sequences in eukaryotic cells.  
     
     
         2 . Use of prokaryotic beta recombinase for transgenic work in eukaryotic cells.  
     
     
         3 . Use according to  claim 1  or  2  for controlling gene expression in eukaryots.  
     
     
         4 . Use according to  claim 1  or  2  for manipulating plant genomes in the generation of transgenic plants.  
     
     
         5 . Use according to  claim 1  or  2  in which the eukaryotic cells are mammalian cells.  
     
     
         6 . Use according to  claim 1  or  2  for manipulating pathogenic and Gram positive bacteria.  
     
     
         7 . Use according to any of the previous claim for site-specific intramolecular recombination between two six sites in eukaryotic cells.  
     
     
         8 . Use according to  claim 7  for promotion of two or more different specific recombination events at a time.  
     
     
         9 . Use according to  claim 7  for mediating exclusively intramolecular reactions.  
     
     
         10 . Use according to  claim 7  in which the prokaryotic beta recombinase promotes the deletion of DNA sequences located between directly oriented six sites in mammalian cells.  
     
     
         11 . Use according to  claim 7  in which the prokaryotic beta recombinase promotes the inversion of DNA sequences located between inverted repeated six sites in mammalian cells.  
     
     
         12 . Use according to  claim 10  in which the prokaryotic beta recombinase promotes deletion of a DNA fragment laying between two directly oriented six sites.  
     
     
         13 . Use according to  claim 12  in which the prokaryotic beta recombinase promotes inversion of a DNA fragment laying between two inversely oriented six sites.  
     
     
         14 . Use according to  claim 13  in which the prokaryotic beta recombinase promotes deletion of a DNA fragment laying between direct repeat specific recognition sequences.  
     
     
         15 . Use according to  claim 13  in which the prokaryotic beta recombinase promotes inversion of a DNA fragment laying between inverted repeated specific recognition sequences.  
     
     
         16 . Use according to  claim 10 ,  11 ,  12  or  13  in which the specific recognition sequence is located as an extrachromosomal DNA substrate.  
     
     
         17 . Use of the gene coding for beta recombinase for catalysing site-specific resolution of DNA sequences in an extrachromosomal target introduced into an eukaryotic cell.  
     
     
         18 . Use of the gene according to  claim 17  for catalysing site-specific resolution of DNA sequences when extrachromosomal target is a plasmid.  
     
     
         19 . Use of the gene according to  claim 17  or  18  in which the introduction is made by transfection.  
     
     
         20 . Use of the gene according to any of  claims 17  to  19  in which the resolution means deletion.  
     
     
         21 . Use of the gene according to any of  claims 17  to  19  in which the resolution means inversion.  
     
     
         22 . Use of the gene according to any of  claims 17  to  21  in which the DNA sequences are allocated between the six sites.  
     
     
         23 . Use of the gene according to any of  claims 17  to  22  when the six sites are integrated in the genome as chromatin associated structures.  
     
     
         24 . Use of the gene according to any of  claims 17  to  22  when the six sites are integrated in the genome and wrapped on a nucleosome, at several locations.  
     
     
         25 . Use according to  claim 1  to develop new techniques for gene delivery in human gene therapy.  
     
     
         26 . Method for development of transgenic animals which includes the use of beta recombinase according to any of claims  1 - 21 .