Novel plasmids and utilization thereof

Abstract

A shuttle vector is constructed by preparing a DNA region replicable in bacteria belonging to the genus Rhodococcus from a Rhodococcus -derived plasmid having the nucleotide sequence set forth as SEQ ID NO: 73 and a plasmid or its DNA fragment having the nucleotide sequence set forth as SEQ ID NO: 74, and a DNA region replicable in E. coli from an E. coli -derived plasmid or its DNA fragment. An aminoketone asymmetric reductase gene is inserted into the shuttle vector, transformants containing the vector are created, and the aminoketone asymmetric reductase and optically active aminoalcohols are produced.

1 - 24 . (canceled) 
     
     
         25 . An isolated DNA fragment having the nucleotide sequence set forth as SEQ ID NO: 77. 
     
     
         26 . An isolated DNA fragment comprising a promoter region having the nucleotide sequence set forth as SEQ ID NO: 77. 
     
     
         27 - 28 . (canceled) 
     
     
         29 . An isolated vector comprising the isolated DNA fragment according to  claim 25 . 
     
     
         30 . The isolated vector according to  claim 29 , having inserted therein an aminoketone asymmetric reductase gene. 
     
     
         31 . The isolated vector according to  claim 30 , wherein the aminoketone asymmetric reductase gene is a nucleic acid coding for a protein comprising the amino acid sequence set forth as SEQ ID NO: 78, or a nucleic acid that codes for a protein having the amino acid sequence set forth as SEQ ID NO: 78 with a deletion, insertion, substitution or addition of one or a plurality of amino acids, and having aminoketone asymmetric reduction activity. 
     
     
         32 . The isolated vector according to  claim 30 , wherein the aminoketone asymmetric reductase gene is a nucleic acid comprising the nucleotide sequence set forth as SEQ ID NO: 79, or a nucleic acid that hybridizes with nucleic acid having a nucleotide sequence complementary to the nucleotide sequence set forth as SEQ ID NO: 79 under stringent conditions, and that codes for a protein having aminoketone asymmetric reduction activity. 
     
     
         33 . A transformant containing the isolated vector according to  claim 29 . 
     
     
         34 . A transformant containing the isolated vector according to  claim 30 . 
     
     
         35 - 38 . (canceled)