PHAGE &phgr;MRU POLYNUCLEOTIDES AND POLYPEPTIDES AND USES THEREOF

Abstract

The invention encompasses phage φmru including phage induction, phage particles, and the phage genome. Also encompassed are phage polypeptides, as well as polynucleotides which encode these polypeptides, expression vectors comprising these polynucleotides, and host cells comprising these vectors. The invention further encompasses compositions and methods for detecting, targeting, permeabilising, and inhibiting microbial cells, especially methanogen cells, using the disclosed phage, polypeptides, polynucleotides, expression vectors, or host cells.

1 - 57 . (canceled) 
     
     
         58 . An isolated polypeptide or peptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1-116, 117, 118, 119, 120-136, and 138-172. 
     
     
         59 . An isolated polypeptide or peptide which comprises: a) an amino acid sequence sharing at least 90% identity with SEQ ID NO:117; b) an amino acid sequence sharing at least 95% identity with SEQ ID NO:118; c) an amino acid sequence sharing at least 90% identity with SEQ ID NO:119; d) an amino acid sequence sharing at least 90% identity with an amino acid sequence selected from the group consisting of SEQ ID NO:1-31, 33-104, 106-112, 114-116, 120-136, and 138-172; e) an amino acid sharing at least 95% identity with an amino acid sequence selected from the group consisting of SEQ ID NO:105 and 113; or f) an amino acid sharing at least 98% identity with amino acid sequence SEQ ID NO:32. 
     
     
         60 . An isolated polypeptide or peptide which comprises: a) at least 15 amino acids of SEQ ID NO:117 or 118; b) at least 15 amino acids of SEQ ID NO:119; c) at least 15 amino acids of an amino acid sequence selected from the group consisting of SEQ ID NO:1-31, 34-73, 75-104, 106-112, 114-116, 120-132, 134-136, and 138-172; d) at least 17 amino acids SEQ ID NO:105; e) at least 19 amino acids of SEQ ID NO:74; or f) at least 22 amino acids of an amino acid sequence selected from the group consisting of SEQ ID NO:33, 113, and 133. 
     
     
         61 . An isolated polynucleotide which comprises: a) a nucleotide sequence which encodes the amino acid sequence SEQ ID NO: 1-116, 117, 118, 119, 120-136, and 138-172; b) a nucleotide sequence selected from the group consisting of SEQ ID NO: 173-341, 342-510, 511, 512, and 513; or e) a nucleotide sequence complementary to (a) or (b). 
     
     
         62 . An isolated polynucleotide which comprises: a) a nucleotide sequence which encodes an amino acid sequence which shares at least 90% identity with SEQ ID NO:117; b) a nucleotide sequence which encodes an amino acid sequence which shares at least 95% identity with SEQ ID NO:118; c) a nucleotide sequence which encodes an amino acid sequence which shares at least 90% identity with SEQ ID NO:119; d) a nucleotide sequence which encodes an amino acid sequence which shares at least 90% identity with an amino acid sequence selected from the group consisting of SEQ ID NO:1-31, 33-104, 106-112, 114-116, 120-136, and 138-172; e) a nucleotide sequence which encodes an amino acid sharing at least 95% identity with an amino acid sequence selected from the group consisting of SEQ ID NO:105 and 113; f) a nucleotide sequence which encodes an amino acid sharing at least 98% identity with amino acid sequence SEQ ID NO:32; or g) a nucleotide sequence complementary to any one of (a) to (f). 
     
     
         63 . An isolated polynucleotide which comprises: a) a nucleotide sequence which encodes at least 15 amino acids of SEQ ID NO:117 or 118; b) a nucleotide sequence which encodes at least 15 amino acids of SEQ ID NO:119; c) a nucleotide sequence which encodes at least 15 amino acids of an amino acid sequence selected from the group consisting of SEQ ID NO:1-31, 34-73, 75-104, 106-112, 114-116, 120-132, 134-136, 138-172; d) a nucleotide sequence which encodes an amino acid sequence which comprises at least 17 amino acids SEQ ID NO:105; e) a nucleotide sequence which encodes an amino acid sequence which comprises at least 19 amino acids of SEQ ID NO:74; f) a nucleotide sequence which encodes an amino acid sequence which comprises at least 22 amino acids of an amino acid sequence selected from the group consisting of SEQ ID NO:33, 113, and 133; or g) a nucleotide sequence complementary to any one of (a) to (f). 
     
     
         64 . A vector which comprises the isolated polynucleotide of any one of  claims 61  to  63 . 
     
     
         65 . A host cell which is genetically modified to express the polypeptide or peptide of any one of  claims 58  to  60 . 
     
     
         66 . A host cell which is genetically modified to comprise the polynucleotide of any one of  claims 61  to  63 . 
     
     
         67 . A host cell which comprises the vector of  claim 64 . 
     
     
         68 . A conjugate molecule or fusion molecule which comprises the polypeptide or peptide of any one of  claims 58  to  60 . 
     
     
         69 . A conjugate molecule or fusion molecule which comprises the polynucleotide of any one of  claims 61  to  63 . 
     
     
         70 . A method of permeabilising a microbial cell comprising: a) optionally, producing or isolating the polypeptide or peptide of any one of  claims 58  to  60 ; and b) contacting the cell with the polypeptide or peptide. 
     
     
         71 . The method of  claim 70 , wherein the cell is a methanogen. 
     
     
         72 . The method of  claim 71 , wherein the cell is  Methanobrevibacter ruminantium.    
     
     
         73 . The method of  claim 72 , wherein the cell is  Methanobrevibacter ruminantium  strain M1 T  (DSM1093). 
     
     
         74 . A method of permeabilising a microbial cell comprising: a) optionally, producing or isolating the conjugate molecule or fusion molecule of  claim 68 ; and b) contacting the cell with the conjugate molecule or the fusion molecule. 
     
     
         75 . The method of  claim 74 , wherein the cell is a methanogen. 
     
     
         76 . The method of  claim 75 , wherein the cell is  Methanobrevibacter ruminantium.    
     
     
         77 . The method of  claim 76 , wherein the cell is  Methanobrevibacter ruminantium  strain MIT (DSM1093).