Human and mammalian DNA replication origin consensus sequences

Abstract

The present invention relates to a human or mammalian DNA replication origin consensus sequence which consists of a sequence selected from the group consisting of CCTMDAWKSGBYTSMAAWYWBCMYTTRSCAAATTCC (SEQ ID NO: 1); and AWMTWAAKRAWRWWKKDAVWWGAKRWWKWVWHRASSACMDWKAAKTWKGGWTWARRYWKGRKMWWTWKAWSDATAKWWWKDAKWKMWRKTT (SEQ ID NO: 4). A method for the control of initiation of mammalian DNA replication which comprises the steps of: a) inserting a consensus sequence coding for a sequence of the present invention together with a DNA fragment to form a vector capable of expression of the DNA fragment; b) introducing the vector of step a) into mammalian cells in vitro.

What is claimed is:  
     
         1 . A human or mammalian DNA replication origin consensus sequence which consists of a sequence selected from the group consisting of: CCTMDAWKSGBYTSMAAWYWBCMYTTRSCAAATTCC (SEQ ID NO: 1), functional variants thereof having a sequence of at least 70% homology, functional fragments thereof of at least 20 nucleotides; AWMTWAAKRAWRWWKKDAVWWGAKRWW KWVWHRASSACMDWKAAKTWKGGWTWARRYWKGRKMWWTWKAWSDATAKWWWKDA KWKMWRKTT (SEQ ID NO: 4), and functional variants thereof having a sequence of at least 70% homology.  
     
     
         2 . A method for the control of initiation of mammalian DNA replication which comprises the steps of: 
 a) inserting a consensus sequence according to  claim 1  together with a DNA fragment to form a vector capable of expression of said DNA fragment;    b) introducing the vector of step a) into mammalian cells in vitro.    
     
     
         3 . The method of  claim 2 , wherein the vector may be selected from the group consisting of viral vectors.  
     
     
         4 . The method of  claim 2 , wherein introducing the vector of step b) is effected by a standard method selected from the group consisting of calcium phosphate co-precipitation transfection, electroporation, micro-injection, liposome-mediated transfection, and virally transmitted DNA.  
     
     
         5 . A DNA sequence for the maintenance of circular plasmid constructs which are capable of being replicated semiconservatively in proliferating mammalian cells, which comprises at least one consensus sequence consisting of a sequence according to  claim 1 .  
     
     
         6 . A DNA sequence suitable for inclusion in mammalian and human artificial chromosome vectors or for gene therapy, which comprises at least one consensus sequence consisting of a sequence according to  claim 1 .  
     
     
         7 . A protein which binds double-stranded DNA, which protein is isolated from HeLa cells and comprises about 150 kDa and two subunits of about 86 and 70 kDa in a glycerol gradient.  
     
     
         8 . An anti-gene to DNA replication, which comprises a doubled-stranded form of a sequence according to  claim 1 .  
     
     
         9 . A method of inhibiting DNA replication in vitro or in vivo, which comprises administering a sequence according to  claim 1  in single-stranded or double-stranded form.  
     
     
         10 . The use of a sequence according to  claim 1  for inhibiting DNA replication in vitro or in vivo, wherein said sequence is in single-stranded or double-stranded form.