DNA Sequences For Regulating Transcription

Abstract

The invention relates to a DNA sequences for regulating transcription of a structural gene encoding a polypeptide in a eukaryotic host cell comprising (a) a first DNA sequence to which RNA polymerase binds which DNA sequence comprises a mRNA initiation site; and further (b) one or more DNA sequence(s) to which RNA polymerase binds with or without a mRNA initiation site. The invention also relates to a DNA construct and an expression vector and a host cell comprising the DNA sequence of the invention.

1 . A DNA sequence for regulating transcription of a structural gene encoding a polypeptide in an  Aspergillus  host cell comprising:
 i) a first DNA sequence comprising nucleotides 7-510 of SEQ ID NO: 1, an RNA polymerase recognition site, and an  Aspergillus nidulans  triose phosphate isomerase leader sequence; and   (ii) one or more additional DNA sequences comprising nucleotides 7-510 of SEQ ID NO:1 with or without an mRNA initiation site,   
       wherein the first DNA sequence and one or more additional DNA sequences are operably linked to the structural gene, and wherein the first DNA sequence and the one or more additional DNA sequences comprise at least 3  Aspergillus  amyR transcription factor binding sites. 
     
     
         2 . The DNA sequence of  claim 1 , wherein the  Aspergillus  host cell is selected from the group consisting of  A. niger,A. oryzae, A. awamori, A. foetidus , or  A. japonicus.    
     
     
         3 . The DNA sequence of  claim 1 , further comprising a third DNA sequence comprising nucleotides 7-510 of SEQ ID NO: 1 with or without a RNA polymerase recognition site and/or mRNA initiation site. 
     
     
         4 . The DNA sequence of  claim 1 , comprising two or more repetitive RNA polymerase binding sites located in tandem or where the RNA polymerase binding sites are closely linked. 
     
     
         5 . A DNA construct comprising a DNA sequence of  claim 1 , operative in an  Aspergillus  host cell and operably linked with a structural gene encoding a polypeptide and a terminator region. 
     
     
         6 . An expression vector comprising a DNA construct of  claim 5 , further comprising a signal peptide-coding region. 
     
     
         7 . An  Aspergillus  host cell comprising a DNA sequence of  claim 4 .