Infectious clones of torque teno virus

Abstract

The present invention is directed to novel nucleotide and amino acid sequences of Torque teno virus (“TTV”), including novel genotypes thereof, all of which are useful in the preparation of vaccines for treating and preventing diseases in swine and other animals. Vaccines provided according to the practice of the invention are effective against multiple swine TTV genotypes and isolates. Diagnostic and therapeutic polyclonal and monoclonal antibodies are also a feature of the present invention, as are infectious clones useful in the propagation of the virus and in the preparation of vaccines. Particularly important aspects of the invention include vaccines that provide TTV ORF1 protein, or peptide fragments thereof, as antigen.

1 . An isolated polynucleotide sequence that comprises a polynucleotide selected from the group consisting of:
 (a 1 ) the DNA of genotype 2 sequence TTV13 (SEQ ID NO: 1); the DNA genotype 2 sequence TTV10 (SEQ ID NO: 2); or a fragment thereof than encodes the TTV capsid protein or a fragment of said protein;   (a 2 ) the DNA of a genotype 1 sequence selected from the group consisting of ttvg1-7 (SEQ ID NO: 4), ttvGT1-17 (SEQ ID NO: 5), ttvGT1-21 (SEQ ID NO: 6), ttvgt1-27 (SEQ ID NO: 3), and ttvgt1-178 (SEQ ID NO: 7) or a fragment thereof than encodes the TTV capsid protein or a fragment of said protein;   (b) the complement of any sequence in (a);   (c) a polynucleotide that hybridizes with a sequence of (a) or (b) under stringent conditions defined as hybriding to filter bound DNA in 0.5M NaHPO 4 , 7% SDS, 1 mM EDTA at 65° C., and washing in 0.1×SSC/0.1% SDS at 68° C.;   (d) a polynucleotide that is at least 70% identical to the polynucleotide of (a) or (b);   (e) a polynucleotide that is at least 80% identical to the polynucleotide of (a) or (b);   (f) a polynucleotide that is at least 90% identical to the polynucleotide of (a) or (b); and   (g) a polynucleotide that is at least 95% identical to the polynucleotide of or (b).   
     
     
         2 . An RNA polynucleotide molecule that is complement of any DNA polynucleotide sequence according to  claim 1 . 
     
     
         3 . A vector or plasmid that comprises a polynucleotide according to  claim 1  or  claim 2 . 
     
     
         4 . A host cell that comprises a vector or plasmid according to  claim 3 . 
     
     
         5 . A virus that be expressed from a nucleotide sequence according to  claim 1 , wherein said virus is live, or fully or partially attenuated. 
     
     
         6 . A vaccine comprising a virus according to  claim 5 . 
     
     
         7 . A DNA vaccine that comprises a polynucleotide sequence of  claim 1 . 
     
     
         8 . A polypeptide encoded by any of the open reading frames of the TTV13 (SEQ ID NO:1) or TTV10 (SEQ ID NO: 2) polynucleotides, or a polypeptide that is at least 90% identical thereto, or to a fragment thereof. 
     
     
         9 . A polypeptide encoded by any of the open reading frames of the ttvg1-7 (SEQ ID NO:10), ttvGT1-17 (SEQ ID NO:11), ttvGT1-21 (SEQ ID NO:12), and ttvgt1-27 (SEQ ID NO:13) polynucleotides, or a polypeptide that is at least 90% identical thereto, or to a fragment thereof. 
     
     
         10 . A vaccine that comprises a polypeptide according to  claim 8  or  claim 9 . 
     
     
         11 . A peptide that comprises: (a) the first 100 N-terminal amino acids of the capsid protein of TTV13 (SEQ NO:1) or TTV10 (SEQ ID NO:2); (b) an amino acid sequence that is at least 90 percent identical theret; or (c) an arginine rich region thereof. 
     
     
         12 . A vaccine composition comprising a peptide of  claim 11 . 
     
     
         13 . A monoclonal or polyclonal antibody composition that binds specifically to a protein or peptide of  claim 8 ,  9  or  11 . 
     
     
         14 . A vaccine that comprises one or more polypeptides encoded by an open reading frame selected from the group consisting of ORF1 ORF2 or ORF3.