Compositions comprising promoter sequences and methods of use

Abstract

Nucleic acid molecules, fragments and variants thereof having promoter activity are provided in the current invention. The invention also provides vectors containing a nucleic acid molecule of the invention and cells comprising the vectors. Methods for making and using the nucleic acid molecules of the invention are further provided.

1 . An isolated nucleic acid molecule having promoter activity, wherein the nucleic acid molecule is selected from the group consisting of:
 (a) a nucleic acid molecule comprising a nucleotide sequence as set forth in any one of SEQ ID NOS:65 or 73;   (b) a nucleic acid molecule that hybridizes to the complement of the nucleic acid of (a) under stringent conditions and has promoter activity, wherein said stringent conditions comprise 6×SSC, 0.1% SDS at about 45° C., followed by a wash of 0.2×SSC and 0.1% SDS at 50-65° C.; and   (c) a nucleic acid molecule having at least 95% identity to the nucleotide sequence set forth in any one of SEQ ID NOS:65 or 73 and has promoter activity.   
     
     
         2 . The nucleic acid molecule of  claim 1  comprising a nucleotide sequence as set forth in any one of SEQ ID NOS:65 or 73. 
     
     
         3 . A recombinant nucleic acid molecule comprising the isolated nucleic acid molecule according to  claim 1  operably associated with a heterologous nucleic acid of interest. 
     
     
         4 . The recombinant nucleic acid molecule according to  claim 3 , wherein said heterologous nucleic acid of interest encodes a protein or peptide. 
     
     
         5 . The recombinant nucleic acid molecule according to  claim 3 , wherein said heterologous nucleic acid of interest encodes an antisense oligonucleotide. 
     
     
         6 . The recombinant nucleic acid molecule according to  claim 3 , wherein said heterologous nucleic acid of interest encodes a ribozyme or interfering RNA. 
     
     
         7 . A plasmid comprising the nucleic acid molecule according to any of  claim 1 . 
     
     
         8 . A method of transforming a cell comprising introducing the nucleic acid molecule according to  claim 1  into a cell. 
     
     
         9 . A cell comprising a heterologous nucleic acid molecule according to  claim 1 . 
     
     
         10 . A cell comprising the plasmid according to  claim 7 . 
     
     
         11 . The cell according to  claim 9 , wherein said cell is a lactic acid producing bacterial cell. 
     
     
         12 . The cell according to  claim 9 , wherein said cell is selected from the group consisting of a cell of a gram positive bacterium, a lactic acid bacteria,  Lactobacillus acidophilus, Lactococcus lactis  or  Lactobacillus gasserei.    
     
     
         13 . A method of controlling the transcription of a nucleic acid of interest in a cell comprising:
 (a) providing or maintaining the cell under non-inducing conditions, said cell comprising the recombinant nucleic acid molecule of  claim 3 , and   (b) subjecting said cell to inducing conditions whereby transcription of said nucleic acid of interest is increased as compared to the level of transcription of said nucleic acid of interest under non-inducing conditions.   
     
     
         14 . The method according to  claim 13 , wherein said nucleic acid of interest encodes a protein or a peptide. 
     
     
         15 . The method according to  claim 13 , wherein said nucleic acid of interest encodes an antisense oligonucleotide or a ribozyme. 
     
     
         16 . The method according to  claim 13 , wherein said inducing conditions described in (b) are produced by an increase or decrease in the concentration of a sugar in said cell relative to the concentration of said sugar in said cell under non-inducing conditions. 
     
     
         17 . The method according to  claim 13 , wherein said inducing conditions described in (b) are produced by the presence of a stress response protein. 
     
     
         18 . A method to express a nucleotide sequence of interest in a cell comprising introducing into said cell the recombinant nucleic acid molecule of  claim 3 . 
     
     
         19 . A method to express a nucleotide sequence of interest in a cell comprising introducing into said cell the nucleic aid molecule of  claim 1 , wherein said nucleic acid molecule is heterologous to said cell and is operably linked to said nucleotide sequence of interest. 
     
     
         20 . An isolated nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:6, wherein said nucleotide sequence has promoter activity. 
     
     
         21 . A method to express a nucleotide sequence of interest in a cell comprising introducing into said cell a recombinant nucleic aid molecule comprising a polynucleotide having at least 95% sequence identity to SEQ ID NO:6 or comprising SEQ ID NO:6, wherein said polynucleotide has promoter activity and is operably linked to the nucleotide sequence of interest.