US2005289666A1PendingUtilityA1

Starch-inducible promoters, recombinant gene constructs, and methods of regulating gene expression

46
Individually held — no corporate assignee on recordPriority: Aug 4, 2000Filed: May 4, 2005Published: Dec 29, 2005
Est. expiryAug 4, 2020(expired)· nominal 20-yr term from priority
Inventors:Brian HookerKeith Douglas MillerJohnway GaoRodney S. SkeenDaniel B. Anderson
C07K 14/39
46
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Claims

Abstract

The invention includes an isolated promoter containing nucleotides 1827 through 2147 of SEQ ID NO.: 1. The invention also includes methods of regulating expression of a gene by fusing the isolated promoter to a coding sequence to form a fused construct, and introducing the fused construct into a host such that the promoter regulates the expression of the gene product within the host. The invention additionally includes a recombinant gene comprising an isolated promoter containing at least nucleotides 1827 through 2147 of SEQ ID NO.: 1, operably linked to a coding region encoding a gene product of interest. The invention includes a host cell containing the recombinant gene.

Claims

exact text as granted — not AI-modified
1 . A method of regulating expression of a gene product comprising: 
 providing a coding region comprising a nucleic acid sequence that encodes a gene product;    fusing the coding region with an isolated promoter to form a fused construct, the isolated promoter comprising nucleotides 1827 through 2147 of SEQ ID NO.: 1; and    introducing the fused construct into a host such that the promoter regulates the expression of the gene product within the host.    
     
     
         2 . The method of  claim 1  wherein the host is a yeast cell, and wherein the introducing comprises integrating the fused construct into genomic DNA.  
     
     
         3 . The method of  claim 2  wherein the yeast is a species other than  Schwanniomyces occidentalis.    
     
     
         4 . The method of  claim 1  wherein the isolated promoter further comprises nucleotides 1297 through 1826 of SEQ ID NO.: 1.  
     
     
         5 . The method of  claim 1  further comprising inducing expression of the gene providing starch to the host.  
     
     
         6 . An isolated gene promoter comprising at least nucleotides 1827 through 2147 of SEQ ID NO.:1.  
     
     
         7 . A vector comprising the isolated promoter of  claim 6 .  
     
     
         8 . The vector of  claim 7  wherein the vector is a plasmid vector.  
     
     
         9 . A recombinant gene comprising: 
 the isolated promoter of  claim 6;  and    a coding region comprising a nucleic acid sequence encoding a gene product other than the  Schwanniomyces occidentals ATCC  26077 glucoamylase gene product, the isolated promoter being operably linked to the coding region.    
     
     
         10 . A host cell comprising the recombinant gene of  claim 9 .  
     
     
         11 . A method of regulating expression of a gene product comprising: 
 providing a coding region comprising a nucleic acid sequence that encodes a gene product;    fusing the coding region with an isolated promoter to form a fused construct, the isolated promoter comprising at least nucleotides 647-1297 of SEQ ID NO.: 1; and    introducing the fused construct into a host such that the promoter regulates the expression of the gene product within the host.    
     
     
         12 . A method of expressing a gene product comprising: 
 providing a starch inducible promoter comprising at least nucleotides 1827 through 2147 of SEQ ID NO.: 1;    operably linking the starch inducible promoter to a coding DNA sequence to form a recombinant gene, the coding DNA sequence encoding a product of interest;    introducing the recombinant gene into a host cell; and    inducing expression of the recombinant gene.    
     
     
         13 . The method of  claim 12  wherein the inducing comprises providing starch into a growth medium.  
     
     
         14 . The method of  claim 13  wherein the starch is the primary carbon source within the growth medium.  
     
     
         15 . The method of  claim 12  wherein the host cell is a yeast.  
     
     
         16 . The method of  claim 12  wherein the host is a plant cell.  
     
     
         17 . The method of  claim 12  wherein the host is a plant protoplast.  
     
     
         18 . The method of  claim 12  wherein the host is a  Nicotiana tabacum  cell.  
     
     
         19 . The method of  claim 12  wherein the gene product is selected from the group consisting of an enzyme, an enzyme cofactor, a ligand, and a receptor.  
     
     
         20 . The method of  claim 12  wherein the gene product is a structural protein.  
     
     
         21 . A host cell comprising a promoter operably linked to a coding sequence which encodes a gene product other than  Schwanniomyces occidentalis  ATCC 26077 glucoamylase gene product, the promoter comprising at least nucleotides 1827 through 2147 of SEQ ID NO.: 1.  
     
     
         22 . The host cell of  claim 21  wherein the host cell is a plant cell.  
     
     
         23 . A method of regulating expression of a gene product comprising: 
 providing a coding region comprising a nucleic acid sequence that encodes a gene product;    fusing the coding region with an isolated promoter to form a fused construct, the isolated promoter comprising nucleotides 486 through 2147 of SEQ ID NO.: 1; and    introducing the fused construct into a host such that the promoter regulates the expression of the gene product within the host.    
     
     
         24 . The method of  claim 23  wherein the host is a yeast.

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