galK

Abstract

The invention provides galK promoter polynucleotides and methods for producing and using such polynucleotides, as well as their variants, agonists and antagonists, and their uses.

What is claimed is:  
     
         1 . An isolated polynucleotide selected from the group consisting of: 
 (i) an isolated polynucleotide comprising a nucleotide sequence which has at least: 
 (a) 70% identity;  
 (b) 80% identity;  
 (c) 90% identity; or  
 (d) 95% identity,  
 to that of SEQ ID NO:1 over the entire length of SEQ ID NO:1;  
   (ii) an isolated polynucleotide which is the polynucleotide of SEQ ID NO:1;    (iii) an isolated polynucleotide obtainable by screening an appropriate library under stingent hybridization conditions with a probe having the sequence of SEQ ID NO:1 or a fragment thereof;    (iv) an isolated polynucleotide comprising a promoter expressibly linked to the galK gene contained in the  Streptococcus pneumoniae ; and    (v) a polynucleotide sequence complementary to said isolated polynucleotide of (i), (ii), (iii) or (iv).    
     
     
         2 . A method for the treatment of an individual: 
 (i) in need of enhanced activity of the polynucleotide of  claim 1  comprising the step of administenng to the individual a therapeutically effective amount of an agonist to said polynucleotide; or    (ii) having need to inhibit activity or expression of the polynucleotide of  claim 1  comprising: 
 (a) administering to the individual a therapeutically effective amount of an antagonist to said polynucleotide; or  
 (b) administering to the individual a therapeutically effective amount of a compound that competes with said polynucleotide for its ligand or binding factor.  
   
     
     
         3 . A method for screening to identify compounds that activate or that inhibit the function of the polynucleotide of  claim 1  which comprises a method selected from the group consisting of: 
 (a) mixing a candidate compound with a solution containing a polynucleotide of  claim 1 , to form a mixture, measuring activity of the polynucleotide in the mixture, and comparing the activity of the mixture to a standard; or  
 (b) detecting the effect of a candidate compound on the production of mRNA or polypeptide by promoter activity of the polynucleotide in cells, using for instance, an ELISA assay, or  
 (c) (1) contacting a composition comprising the polynucleotide with the compound to be screened under conditions to permit interaction between the compound and the polynucleotide to assess the interaction of a compound, such interaction being associated with a second component capable of providing a detectable signal in response to the interaction of the polynucleotide with the compound; and  
 (2) determining whether the compound interacts with and activates or inhibits an activity of the polynucleotide by detecting the presence or absence of a signal generated from the interaction of the compound with the polynucleotide.  
 
     
     
         4 . An agonist or an antagonist of the activity of or expression from polynucleotide of  claim 1 .  
     
     
         5 . A host cell comprising the polynucleotide of  claim 1 .  
     
     
         6 . A polynucleotide consisting of a polynucleotide of the formula: 
       X—(R 1 ) m —(R 2 )—(R 3 ) n —Y 
       wherein, at the 5′ end of the molecule, X is hydrogen, a metal or a modified nucleotide residue, or together with Y defines a covalent bond, and at the 3′ end of the molecule, Y is hydrogen, a metal, or a modified nucleotide residue, or together with X defines the covalent bond, each occurrence of R 1  and R 3  is independently any nucleic acid residue or modified nucleic acid residue, m is an integer between 1 and 3000 or zero, n is an integer between 1 and 3000 or zero, and R 2  is a nucleic acid sequence or modified nucleic acid sequence set forth in SEQ ID NO:1.  
     
     
         7 . The method of ascertaining the functionality or essentiality of the target gene (gene-of-interest) in a cell comprising the transcription or expression of gene-of-interest by expressing an anti-sense sequence to the gene-of-interest under the transcriptional control of the promoter polynucleotides of  claim 1 .  
     
     
         8 . The method of ascertaining the functionality or essentiality of the target gene (gene-of interest), in a cell, comprising: 
 (a) disabling (“knocking-out”) the gene-of-interest;    (b) re-introducing, at the target gene locus, the gene-of-interest now under the operational control of the inducible promoter polynucleotides of claim  1 ; and    (c) adding the inducer thereby providing information to the essentiality or functionality of the gene of interest.    
     
     
         9 . A method of determining a functionality or essentiality of the target gene (gene-of-interest), in a cell, comprising: 
 (a) replacing the promoter sequence of the gene-of-interest by the promoter polynucleotide of claim  1 ; and    (b) adding the repressor thereby providing information to the essentiality or functionality of the gene-of-interest.    
     
     
         10 . A method of determining a protein target of a antibacterial compound comprising: 
 (a) placing the protein target under the control of the promoter polynucleotide of claim  1 ; and    (b) adding the inducer or repressor thereby changing accordingly the susceptibility of the bacteria to the compound.