Enterococcus faecalis polynucleotides and polypeptides

Abstract

The present invention relates to novel genes from E. faecalis and the polypeptides they encode. Also provided as are vectors, host cells, antibodies and methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of E. faecalis polypeptide activity. The invention additionally relates to diagnostic methods for detecting Enterococcus nucleic acids, polypeptides and antibodies in a biological sample. The present invention further relates to novel vaccines for the prevention or attenuation of infection by Enterococcus.

1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding any one of the amino acid sequences of the polypeptides shown in Table 1; or    (b) a nucleotide sequence complementary to any one of the nucleotide sequences in (a).    (c) a nucleotide sequence at least 95% identical to any one of the nucleotide sequences shown in Table 1; or,    (d) a nucleotide sequence at least 95% identical to a nucleotide sequence complementary to any one of the nucleotide sequences shown in Table 1.    
     
     
         2 . An isolated nucleic acid molecule of  claim 1  comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a) or (b) of  claim 1 .  
     
     
         3 . An isolated nucleic acid molecule of  claim 1  comprising a polynucleotide which encodes an epitope-bearing portion of a polypeptide in (a) of  claim 1 .  
     
     
         4 . The isolated nucleic acid molecule of  claim 3 , wherein said epitope-bearing portion of a polypeptide comprises an amino acid sequence listed in Table 4.  
     
     
         5 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector.  
     
     
         6 . A recombinant vector produced by the method of  claim 5 .  
     
     
         7 . A host cell comprising the vector of  claim 6 .  
     
     
         8 . A method of producing a polypeptide comprising: 
 (a) growing the host cell of  claim 7  such that the protein is expressed by the cell; and    (b) recovering the expressed polypeptide.    
     
     
         9 . An isolated polypeptide comprising a polypeptide selected from the group consisting of: 
 (a) a polypeptide consisting of one of the complete amino acid sequences of Table 1;    (b) a polypeptide consisting of one the complete amino acid sequences of Table 1 except the N-terminal residue;    (c) a fragment of the polypeptide of (a) having biological activity; and    (d) a fragment of the polypeptide of (a) which binds to an antibody specific for the polypeptide of (a).    
     
     
         10 . An isolated antibody specific for the polypeptide of  claim 9 .  
     
     
         11 . A polypeptide produced according to the method of  claim 8 .  
     
     
         12 . An isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of an amino acid sequence of any one of the polypeptides in Table 1.  
     
     
         13 . An isolated polypeptide antigen comprising an amino acid sequence of an  E. faecalis  epitope shown in Table 4.  
     
     
         14 . An isolated nucleic acid molecule comprising a polynucleotide with a nucleotide sequence encoding a polypeptide of  claim 9 .  
     
     
         15 . A hybridoma which produces an antibody of  claim 10 .  
     
     
         16 . A vaccine, comprising: 
 (1) one or more  E. faecalis  polypeptides selected from the group consisting of a polypeptide of  claim 9;  and    (2) a pharmaceutically acceptable diluent, carrier, or excipient;    wherein said polypeptide is present, in an amount effective to elicit protective antibodies in an animal to a member of the  Enterococcus  genus.    
     
     
         17 . A method of preventing or attenuating an infection caused by a member of the  Enterococcus  genus in an animal, comprising administering to said animal a polypeptide of  claim 9 , wherein said polypeptide is administered in an amount effective to prevent or attenuate said infection.  
     
     
         18 . A method of detecting  Enterococcus  nucleic acids in a biological sample comprising: 
 (a) contacting the sample with one or more nucleic acids of  claim 1 , under conditions such that hybridization occurs, and    (b) detecting hybridization of said nucleic acids to the one or more  Enterococcus  nucleic acid sequences present in the biological sample.    
     
     
         19 . A method of detecting  Enterococcus  nucleic acids in a biological sample obtained from an animal, comprising: 
 (a) amplifying one or more  Enterococcus  nucleic acid sequences in said sample using polymerase chain reaction, and    (b) detecting said amplified  Enterococcus  nucleic acid.    
     
     
         20 . A kit for detecting  Enterococcus  antibodies in a biological sample obtained from an animal, comprising 
 (a) a polypeptide of  claim 9  attached to a solid support; and    (b) detecting means.    
     
     
         21 . A method of detecting  Enterococcus  antibodies in a biological sample obtained from an animal, comprising 
 (a) contacting the sample with a polypeptide of  claim 9;  and    (b) detecting antibody-antigen complexes.