Methods and compositions involving promoters derived from yarrowia lipolytica

Abstract

The current methods and compositions provide for nucleotide sequences of promoters from Yarrowia lipolytica which may be used to drive gene expression in a cell. In some aspects, the promoters are useful for modulating lipid production in oleaginous organisms such as yeast.

1 - 42 . (canceled) 
     
     
         43 . A method of producing a lipid, the method comprising culturing a recombinant cell, thereby producing the lipid, wherein the recombinant cell comprises a nucleic acid molecule comprising a promoter sequence having at least 80% sequence identity to SEQ ID NO: 1, 2, 5, 6, or 7, wherein the promoter sequence is operably linked to a coding sequence, wherein the coding sequence is heterologous to the promoter sequence. 
     
     
         44 . The method of  claim 43 , wherein the coding sequence comprises SCT1. 
     
     
         45 . The method of  claim 43 , wherein the recombinant cell is a yeast cell. 
     
     
         46 . The method of  claim 43 , wherein the recombinant cell is an Arxula,  Aspergillus, Aurantiochytrium, Candida, Claviceps, Cryptococcus, Cunninghamella, Geotrichum, Hansenula, Kluyveromyces, Kodamaea, Leucosporidiella, Lipomyces, Mortierella, Ogataea, Pichia, Prototheca, Rhizopus, Rhodosporidium, Rhodotorula, Saccharomyces, Schizosaccharomyces, Tremella, Trichosporon, Wickerhamomyces , or  Yarrowia  cell. 
     
     
         47 . The method of  claim 46 , wherein the recombinant cell is an Arxula adeninivorans,  Aspergillus niger, Aspergillus orzyae, Aspergillus terreus, Aurantiochytrium limacinum, Candida utilis, Claviceps purpurea, Cryptococcus albidus, Cryptococcus curvatus, Cryptococcus ramirezgomezianus, Cryptococcus terreus, Cryptococcus wieringae, Cunninghamella echinulata, Cunninghamella japonica, Geotrichum fermentans, Hansenula polymorpha, Kluyveromyces lactis, Kluyveromyces marxianus, Kodamaea ohmeri, Leucosporidiella creatinivora, Lipomyces lipofer, Lipomyces starkeyi, Lipomyces tetrasporus, Mortierella isabellina, Mortierella alpina, Ogataea polymorpha, Pichia ciferrii, Pichia guilliermondii, Pichia pastoris, Pichia stipites, Prototheca zopfii, Rhizopus arrhizus, Rhodosporidium babjevae, Rhodosporidium toruloides, Rhodosporidium paludigenum, Rhodotorula glutinis, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Tremella enchepala, Trichosporon cutaneum, Trichosporon fermentans, Wickerhamomyces ciferrii  or a  Yarrowia lipolytica  cell. 
     
     
         48 . The method of  claim 47 , wherein the recombinant cell is a  Yarrowia lipolytica.    
     
     
         49 . The method of  claim 43 , wherein the recombinant cell is cultured under conditions sufficient to express the coding sequence. 
     
     
         50 . The method of  claim 43 , wherein the recombinant cell is cultured for at least 10 passages. 
     
     
         51 . The method of  claim 43 , wherein the promoter sequence is capable of inducing increased expression of the coding sequence in a growth phase of a recombinant cell relative to a lipid accumulation phase of the recombinant cell. 
     
     
         52 . The method of  claim 43 , wherein the promoter sequence has at least 85% sequence identity to SEQ ID NO:1, 2, 5, 6 or 7. 
     
     
         53 . The method of  claim 43 , wherein the promoter sequence has at least 90% sequence identity to SEQ ID NO:1, 2, 5, 6 or 7. 
     
     
         54 . The method of  claim 43 , wherein the promoter sequence has at least 95% sequence identity to SEQ ID NO:1, 2, 5, 6 or 7. 
     
     
         55 . The method of  claim 43 , wherein the promoter sequence comprises SEQ ID NO:1, 2, 5, 6 or 7. 
     
     
         56 . The method of  claim 43 , further comprising extracting the lipid from the recombinant cell, thereby producing a lipid product. 
     
     
         57 . The method of  claim 43 , wherein the lipid is a fatty acid, further comprising extracting the fatty acid from the recombinant cell, thereby producing a fatty acid product; and esterifying the fatty acid product to a fatty acid ester product.