Lactobacillus rhamnosus polynucleotides, polypeptides and methods for using them

Abstract

Novel polynucleotides isolated from Lactobacillus rhamnosus , as well as probes and primers, genetic constructs comprising the polynucleotides, biological materials, including plants, microorganisms and multicellular organisms incorporating the polynucleotides, polypeptides expressed by the polynucleotides, and methods for using the polynucleotides and polypeptides are disclosed.

1 . An isolated polynucleotide comprising a sequence selected from the group consisting of: SEQ ID NO: 1-33.  
     
     
         2 . An isolated polynucleotide comprising a sequence selected from the group consisting of: 
 (a) complements of SEQ ID NO: 1-33;    (b) reverse complements of SEQ ID NO: 1-33; and    (c) reverse sequences of SEQ ID NO: 1-33.    
     
     
         3 . An isolated polynucleotide comprising a sequence selected from the group consisting of: 
 (a) sequences having at least 75%, identity to a sequence of SEQ ID NO: 1-33;    (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 1-33; and    (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 1-33, wherein the polynucleotide encodes a polypeptide having substantially the same functional properties as a polypeptide encoded by SEQ ID NO: 1-33.    
     
     
         4 . An isolated polynucleotide comprising a sequence selected from the group consisting of: 
 (a) nucleotide sequences that are 200-mers of a sequence recited in SEQ ID NO: 1-33;    (b) nucleotide sequences that are 100-mers of a sequence recited in SEQ ID NO: 1-33;    (c) nucleotide sequences that are 40-mers of a sequence recited in SEQ ID NO: 1-33; and    (d) nucleotide sequences that are 20-mers of a sequence recited in SEQ ID NO: 1-33;    
     
     
         5 . An isolated oligonucleotide probe or primer comprising at least 10 contiguous residues complementary to 10 contiguous residues of a nucleotide sequence recited in any one of claims  1 - 3 .  
     
     
         6 . A kit comprising a plurality of oligonucleotide probes or primers of  claim 5 .  
     
     
         7 . A genetic construct comprising a polynucleotide of any one of claims  1 - 3 .  
     
     
         8 . A transgenic host cell comprising a genetic construct according to  claim 7 .  
     
     
         9 . A transgenic non-human organism comprising a transgenic host cell of  claim 8 .  
     
     
         10 . The transgenic organism of  claim 9 , wherein the organism is selected from the group consisting of  Lactobacillus  species.  
     
     
         11 . An isolated polynucleotide comprising a nucleotide sequence that differs from a nucleotide sequence recited in SEQ ID NO: 1-33 as a result of deletions and/or insertions totalling less than 15% of the total sequence length.  
     
     
         12 . The isolated polynucleotide of  claim 11 , wherein the nucleotide sequence differs from a nucleotide sequence recited in SEQ ID NO: 1-33 as a result of substitutions, insertions, and/or deletions totalling less than 10% of the total sequence length.  
     
     
         13 . A genetic construct comprising, in the 5′-3′ direction: 
 (a) a gene promoter sequence; and    (b) a polynucleotide sequence comprising at least one of the following: (1) a polynucleotide coding for at least a functional portion of a polypeptide of SEQ ID NO: 42-75; and (2) a polynucleotide comprising a non-coding region of a polynucleotide of any one of claims  1 - 3 .    
     
     
         14 . The genetic construct of  claim 13 , wherein the polynucleotide is in a sense orientation.  
     
     
         15 . The genetic construct of  claim 13 , wherein the polynucleotide is in an anti-sense orientation.  
     
     
         16 . The genetic construct of  claim 13 , wherein the gene promoter sequence is functional in a prokaryote or eukaryote.  
     
     
         17 . A transgenic host cell comprising a construct of  claim 13 .  
     
     
         18 . A transgenic organism comprising a transgenic host cell according to  claim 17 , or progeny thereof.  
     
     
         19 . The transgenic organism of  claim 18 , wherein the organism is selected from the group consisting of  Lactobacillus  species.  
     
     
         20 . A method for modulating the activity of a polypeptide in an organism, comprising stably incorporating into the genome of the organism a polynucleotide of any one of claims  1 - 3 .  
     
     
         21 . The method of  claim 20 , wherein the organism is a microbe.  
     
     
         22 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 42-75.  
     
     
         23 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: 
 (a) sequences having at least 75% identity to a sequence of SEQ ID NO: 42-75;    (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 42-75; and    (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 42-75, wherein the polypeptide has substantially the same functional properties as a polypeptide of SEQ ID NO: 42-75.    
     
     
         24 . An isolated polypeptide encoded by a polynucleotide of any one of claims  1 - 3 .  
     
     
         25 . An isolated polynucleotide that encodes a polypeptide of any one of claims  22  and  23 .  
     
     
         26 . A fusion protein comprising at least one polypeptide according to any one of claims  22  and  23 .  
     
     
         27 . A composition comprising a polypeptide according to any one of claims  22  and  23  and at least one component selected from the group consisting of: physiologically acceptable carriers and immunostimulants.  
     
     
         28 . A composition comprising a polynucleotide according to any one of claims  1 - 3  and at least one component selected from the group consisting of: physiologically acceptable carriers and immunostimulants.  
     
     
         29 . A method for treating a disorder in a mammal, comprising administering a composition according to  claim 27 .  
     
     
         30 . A method for treating a disorder in a mammal, comprising administering a composition according to  claim 28 .  
     
     
         31 . A method for modifying a property of a microbe, comprising modulating the polynucleotide content or composition of the microbe by transforming the microbe with a polynucleotide of any one of claims  1 - 3 .  
     
     
         32 . The method of  claim 31 , wherein the microbe is used in the manufacture of a milk-derived product, food product, food additive, nutritional supplement, bioactive substance or probiotic supplement.  
     
     
         33 . A method for modifying at least one property of a product, food, food additive, nutritional supplement or probiotic supplement, wherein the product, food, food additive, nutritional supplement or probiotic supplement is prepared from milk, the method comprising adding a polypeptide of any one of claims  22 - 24  to the milk.  
     
     
         34 . The method of  claim 33 , wherein the property is selected from the group consisting of: flavor; aroma; texture; nutritional benefits; immune system modulation; and health benefits.  
     
     
         35 . A food product comprising an isolated polypeptide of any one of claims  22  and  23 .  
     
     
         36 . The food product of  claim 35 , wherein the food product is derived from milk.  
     
     
         37 . The food product of  claim 36 , wherein the food product is selected from the group consisting of: cheese; and yoghurt.  
     
     
         38 . The food product of  claim 35 , wherein the food product has at least one modified property selected from the group consisting of: flavor; aroma; texture; nutritional benefits; immune system modulation; and health benefits.