Ketogulonigenium endogenous plasmids

Abstract

The present invention relates, in general, to a novel genus of bacteria known as Ketogulonigenium. The present invention further relates to transformed Ketogulonigenium, and methods of transforming Ketogulonigenium. The present invention also relates to nucleic acid molecules, and vectors.

What is claimed is:  
     
         1 . Ketogulonigenium comprising a transgene comprising a DNA sequence from an endogenous Ketogulonigenium plasmid.  
     
     
         2 . The Ketogulonigenium of  claim 1 , wherein said endogenous Ketogulonigenium plasmid is contained in Deposit Number NRRL B-21627.  
     
     
         3 . A method for producing the Ketogulonigenium of  claim 1 , comprising: 
 (a) transforming a Ketogulonigenium strain with a transgene comprising a DNA sequence from an endogenous Ketogulonigenium plasmid; and    (b) obtaining a stably transformed strain of Ketogulonigenium.    
     
     
         4 . The method of  claim 3 , wherein said transforming comprises conjugation.  
     
     
         5 . The method of  claim 3 , wherein said transforming comprises electroporation.  
     
     
         6 . The method of  claim 3 , wherein said DNA sequence is derived from an endogenous plasmid contained in Deposit Number NRRL B-21627.  
     
     
         7 . A method for conjugative transfer of a vector from  E. coli  to Ketogulonigenium comprising culturing said  E. coli  with said Ketogulonigenium under such conditions wherein said  E. coli  transfers said vector to said Ketogulonigenium.  
     
     
         8 . The method of  claim 7 , wherein said vector comprises pDELIA8.  
     
     
         9 . A method for transforming Ketogulonigenium comprising the process of inserting a vector into said Ketogulonigenium, wherein said insertion comprises electroporation, wherein said vector comprises pMF1014-α.  
     
     
         10 . An isolated or purified nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of 
 the nucleotide sequence in SEQ ID NO:1,    the nucleotide sequence in SEQ ID NO:2,    the nucleotide sequence in SEQ ID NO:3,    the nucleotide sequence in SEQ ID NO:4,    a nucleotide sequence of the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30418,    a nucleotide sequence of the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30419,    a nucleotide sequence of the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30435,    a nucleotide sequence complementary to SEQ ID NO:1,    a nucleotide sequence complementary to SEQ ID NO:2,    a nucleotide sequence complementary to SEQ ID NO:3,    a nucleotide sequence complementary to SEQ ID NO:4,    a nucleotide sequence complementary to the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30418,    a nucleotide sequence complementary to the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30419, and    a nucleotide sequence complementary to the Ketogulonigenium portion of the plasmid contained in NRRL Deposit No. B-30435.    
     
     
         11 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to said nucleotide sequence of  claim 10 , wherein said polynucleotide which hybridizes, does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         12 . A vector comprising the molecule of  claim 10  and at least one marker gene.  
     
     
         13 . A vector comprising: 
 (a) the nucleic acid molecule of claim  10 ;    (b) a terminator of transcription;    (c) a promoter; and    (d) a discrete series of restriction endonuclease recognition sites, said series being between said promoter and said terminator.    
     
     
         14 . An isolated or purified nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a nucleotide sequence of a Ketogulonigenium plasmid replicon found on any one of the endogenous plasmids contained in Deposit No. NRRL B-21627.  
     
     
         15 . The isolated or purified nucleic acid molecule of  claim 14  comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of 
 the nucleotide sequence in SEQ ID NO:5,  
 the nucleotide sequence in SEQ ID NO:6,  
 the nucleotide sequence in SEQ ID NO:7,  
 a nucleotide sequence complementary to SEQ ID NO:5,  
 a nucleotide sequence complementary to SEQ ID NO:6, and  
 a nucleotide sequence complementary to SEQ ID NO:7.  
 
     
     
         16 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to said nucleotide sequence of  claim 14 , wherein said polynucleotide which hybridizes, does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues.  
     
     
         17 . An isolated or purified vector comprising the nucleic acid molecule of  claim 14 .  
     
     
         18 . The vector of  claim 16 , comprising a replicon functional in  E. Coli.    
     
     
         19 . The vector of  claim 16 , comprising a replicon functional in an organism selected from the genera comprising Acetobacter, Corynebacterium, Rhodobacter, Paracoccus, Roseobacter, Pseudomonas, Pseudo gluconobacter, Gluconobacter, Serratia, Mycobacterium, Streptomyces and Bacillus.  
     
     
         20 . A transformed cell of  E. coli  comprising the vector of  claim 17 .  
     
     
         21 . A transformed cell of the genus Ketogulonigenium comprising the vector of  claim 17 .  
     
     
         22 . The vector of  claim 17 , wherein said Ketogulonigenium replicon is selected from the group comprising the nucleotide sequences of SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7.