Protein production in microorganisms of the phylum labyrinthulomycota

Abstract

The present invention relates to recombinant cells and microorganisms of the phylum Labyrinthulomycota and their use in heterologous protein production. Novel promoter, terminator, and signal sequences for efficient production and, optionally, secretion of polypeptides from recombinant host cells and microorganisms are also encompassed by the present invention.

1 - 28 . (canceled) 
     
     
         29 . A recombinant nucleic acid molecule comprising:
 (a) a polynucleotide sequence having at least 90% sequence identity to the polynucleotide sequence of SEQ ID NO: 3; or   (b) a polynucleotide sequence that is fully complementary to the polynucleotide sequence of (a).   
     
     
         30 . The recombinant nucleic acid molecule of  claim 29 , wherein the polynucleotide sequence has at least 95% sequence identity to the polynucleotide sequence of SEQ ID NO: 3. 
     
     
         31 . The recombinant nucleic acid molecule of  claim 29 , wherein the polynucleotide sequence is SEQ ID NO: 3. 
     
     
         32 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over a sequence of at least 10 nucleotides of SEQ ID NO: 3. 
     
     
         33 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over a sequence of at least 20 nucleotides of SEQ ID NO: 3. 
     
     
         34 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over a sequence of at least 50 nucleotides of SEQ ID NO: 3. 
     
     
         35 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over a sequence of at least 100 nucleotides of SEQ ID NO: 3. 
     
     
         36 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over a sequence of at least 300 nucleotides of SEQ ID NO: 3. 
     
     
         37 . The recombinant nucleic acid molecule of  claim 29  or  claim 30 , wherein the % sequence identity can be found over the entire length of SEQ ID NO: 3. 
     
     
         38 . The recombinant nucleic acid molecule of  claim 29 , wherein the recombinant nucleic acid molecule is a vector. 
     
     
         39 . The recombinant nucleic acid molecule of  claim 29 , wherein the polynucleotide sequence is operably linked to a polynucleotide sequence encoding a protein. 
     
     
         40 . The recombinant nucleic acid molecule of  claim 39 , wherein the protein is operably linked to a secretion signal. 
     
     
         41 . A host cell comprising the recombinant nucleic acid molecule of  claim 29 . 
     
     
         42 . The host cell of  claim 41 , wherein the host cell is a member of the order  Thraustochytriales.    
     
     
         43 . The host cell of  claim 42 , wherein the host cell is a  Schizochytrium  or a  Thraustochytrium.    
     
     
         44 . A method for production of a protein encoded by a polynucleotide that is operably linked to the polynucleotide of  claim 29 , comprising:
 (a) culturing a recombinant microorganism of the order Thraustochytriales in a medium, wherein the recombinant microorganism comprises recombinant nucleic acid molecule of  claim 39  or  claim 40 ; and   (b) producing the protein made in step (a).   
     
     
         45 . The method of  claim 44 , wherein the protein is recovered from an isolated Thraustochytriales biomass. 
     
     
         46 . The method of  claim 44 , wherein the protein accumulates in the microorganism. 
     
     
         47 . The method of  claim 44 , wherein the protein accumulates in a membrane of the microorganism. 
     
     
         48 . The method of  claim 44 , wherein the protein is recovered from the culture medium. 
     
     
         49 . The method of  claim 44 , wherein the protein is secreted. 
     
     
         50 . The recombinant nucleic acid molecule of  claim 38 , wherein said vector is deposited under ATCC Accession No. PTA-9616.