Virulence genes, proteins, and their use

Abstract

A series of genes from Streptococcus pyogenes are shown to encode products which are implicated in virulence. The identification of these genes therefore allows attuated microorganisms to be produced. Furthermore, the genes or their encoded products can be used in the manufacture of vaccines for therapeutic application.

1 . A peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.  
     
     
         2 . The peptide according to  claim 1 , wherein the sequence similarity or identity is at least 60%.  
     
     
         3 . The peptide according to  claim 1 , wherein the sequence similarity or identity is at least 90%.  
     
     
         4 . The peptide according to  claim 1 , comprising an amino acid sequence selected from the group consisting of SEQ ID NOS. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 and 62.  
     
     
         5 . A polynucleotide encoding a peptide wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity, and functional fragments thereof.  
     
     
         6 . A host transformed to express a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.  
     
     
         7 . An attenuated microorganism comprising a mutation that disrupts the expression of a polynucleotide encoding a peptide, wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.  
     
     
         8 . The microorganism according to  claim 7 , wherein the mutation is insertional inactivation or a gene deletion.  
     
     
         9 . The microorganism according to  claim 7 , wherein the microorganism is a Group A  Streptococcus.    
     
     
         10 . The microorganism according to  claim 7 , comprising a mutation in a further nucleotide sequence.  
     
     
         11 . The microorganism according to  claim 7 , comprising a heterologous antigen, therapeutic peptide or nucleic acid.  
     
     
         12 . A vaccine comprising a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof; or the means for expressing said peptide.  
     
     
         13 . A vaccine comprising an attenuated microorganism comprising a mutation that disrupts the expression of a polynucleotide encoding a peptide wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and fuictional fragments thereof.  
     
     
         14 . A method for the treatment or prevention of a condition associated with infection by  Streptococcal  or Gram-positive bacteria wherein said method comprises administering to a patient in need of such treatment or prevention one or more of the group consisting of: 
 (i) a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof;    (ii) a polynucleotide encoding a peptide wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity, and functional fragments thereof;    (iii) a host transformed to express a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof;    (iv) an attenuated microorganism comprising a mutation that disrupts the expression of a polynucleotide encoding a peptide, wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof;    (v) a vaccine comprising a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof; or the means for expressing said peptide; and    (vi) a vaccine comprising an attenuated microorganism comprising a mutation that disrupts the expression of a polynucleotide encoding a peptide wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.    
     
     
         15 . The method according to  claim 14 , wherein the condition is pneumonia.  
     
     
         16 . The method according to  claim 14 , for veterinary treatment.  
     
     
         17 . An antibody raised against a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.  
     
     
         18 . A screening assay for the identification of an antimicrobial drug wherein said screening assay utilizes one or more of the group consisting of: 
 (i) a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof;    (ii) a polynucleotide encoding a peptide wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity, and functional fragments thereof,    (iii) a host transformed to express a peptide encoded by an operon comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof, and    (iv) an attenuated microorganism comprising a mutation that disrupts the expression of a polynucleotide encoding a peptide, wherein said polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59 and 61 of  S. pyogenes,  related sequences obtainable from Gram-positive bacterium and having at least 40% sequence similarity or identity at the peptide or nucleotide level, and functional fragments thereof.