Nucleotide sequences coding for the lysRI gene

Abstract

The present invention relates to polynucleotides corresponding to the lysR1 gene and which encode a LysR1 transcriptional regulator, methods of producing L-amino acids, and methods of screening for polynucleotides which encode proteins having LysR3 transcriptional regulator activity.

1 - 22 . (canceled)  
     
     
         23 . A process for producing L-amino acids comprising culturing a recombinant bacterial cell in a medium suitable for producing L-amino acids, and collecting the L-amino acids produced wherein said bacterial cell comprises an attenuated lysR1 gene.  
     
     
         24 . The process of  claim 23 , wherein said recombinant bacterial cell is a  Coryneform bacterium  or  Brevibacterim.    
     
     
         25 . The process of  claim 24 , wherein said recombinant bacterial cell is selected from the group consisting of  Coryneform glutamicum, Corynebacterium acetoglutamicum, Corynebacterium acetoacidophilum, Corynebacterium melassecola, Corynebacterium thermoaminogenes, Brevibacteriumflavum, Brevibacterium lactofermentum, Brevibacterium divaricatum.    
     
     
         26 . The process of  claim 23 , wherein said lysR1 gene prior to being attenuated comprises the polynucleotide sequence of SEQ ID NO: 1 or a polynucleotide which hybridizes under stringent conditions to SEQ ID NO: 1 or the full-complement of SEQ ID NO: 1 wherein the stringent conditions comprise washing in 0.5×SSC at a temperature of 68° C., and wherein the polynucleotide encodes a protein that inhibits lysine production in a bacterial cell.  
     
     
         27 . The process of  claim 23 , wherein said L-amino acid is L-lysine.  
     
     
         28 . The process of  claim 23 , wherein said L-amino acid is L-valine.  
     
     
         29 . The process of  claim 23 , wherein said recombinant bacterial cell further comprises at least one gene whose expression is enhanced, wherein said gene is selected from the group consisting of dapA, eno, zwf, pyc, and lysE.  
     
     
         30 . The process of  claim 23 , wherein said recombinant bacterial cell further comprises at least one gene whose expression is attenuated, wherein said gene is selected from the group consisting of pck, pgi, and poxB.  
     
     
         31 . A process for screening for polynucleotides which encode a protein having LysR1 transcriptional regulatory activity comprising hybridizing the isolated polynucleotide which encodes a protein comprising the amino acid sequence of SEQ ID NO:2 to the polynucleotide to be screened; expressing the polynucleotide to produce a protein; and detecting the presence or absence of LysR1 transcriptional regulatory activity in said protein.  
     
     
         32 . A process for screening for polynucleotides which encode a protein having LysR1 transcriptional regulatory activity comprising hybridizing isolated polynucleotide comprising SEQ ID NO: 1 to the polynucleotide to be screened; expressing the polynucleotide to produce a protein; and detecting the presence or absence of LysR1 transcriptional regulatory activity in said protein.  
     
     
         33 . A method for detecting a nucleic acid with at least 70% homology to an isolated polynucleotide which encodes a protein comprising SEQ ID NO:2, comprising contacting a nucleic acid sample with a probe or primer comprising at least 15 consecutive nucleotides of said isolated polvnucleotide, or at least 15 consecutive nucleotides of the complement thereof.  
     
     
         34 . A method for producing a nucleic acid with at least 70% homology to an isolated polynucleotide which encodes a protein comprising SEQ ID NO:2, comprising contacting a nucleic acid sample with a primer comprising at least 15 consecutive nucleotides of the isolated polynucleotide, or at least 15 consecutive nucleotides of the complement thereof.  
     
     
         35 . A method for detecting a nucleic acid with at least 70% homology to isolated polynucleotide comprising SEQ ID NO: 1, comprising contacting a nucleic acid sample with a probe or primer comprising at least 15 consecutive nucleotides of the nucleotide sequence of said isolated polynucleotide, or at least 15 consecutive nucleotides of the complement thereof.  
     
     
         36 . A method for producing a nucleic acid with at least 70% homology to comprising contacting a nucleic acid sample with a primer comprising at least 15 consecutive nucleotides of the isolated polynucleotide, or at least 15 consecutive nucleotides of the complement thereof.  
     
     
         37 - 38 . (canceled)  
     
     
         39 . An isolated polypeptide comprising the amino acid sequence of SEQ ID NO:2.