Nannochloropsis spliced leader sequences and uses therefor

Abstract

The present invention relates to the culture and manipulation of microorganisms for biotech applications, and is based on the discovery and characterization of spliced leader sequences identified in transcripts from Nannochloropsis species. In particular, the invention provides nucleic acid compositions comprising a SL sequence operably linked to a protein-encoding gene. Further provided are compositions and methods for enhanced gene expression in recombinant microorganisms as well as methods for identification and/or isolation of nucleic acid molecules tagged with a spliced leader sequence.

What is claimed is: 
     
         1 . A nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, and a full length complement of any thereof; further wherein the nucleic acid sequence is operably linked to a heterologous regulatory element. 
     
     
         2 . The nucleic acid molecule according to  claim 1 , wherein the nucleic acid sequence has a nucleic acid sequence selected from the group consisting of SEQ ID NO:2, and SEQ ID NO:3. 
     
     
         3 . The nucleic acid molecule according to  claim 1 , wherein the heterologous regulatory element is a promoter or a terminator. 
     
     
         4 . The nucleic acid molecule according to  claim 3 , wherein the regulatory element is a promoter. 
     
     
         5 . The nucleic acid molecule according to  claim 1 , further comprising a coding sequence for a polypeptide downstream of the nucleic acid sequence. 
     
     
         6 . The nucleic acid sequence according to  claim 1 , further comprising a restriction endonuclease site. 
     
     
         7 . The nucleic acid molecule according to  claim 5 , wherein the coding sequence for a polypeptide is not from a  Nannochloropsis  species. 
     
     
         8 . A recombinant host cell comprising a nucleic acid molecule according to  claim 1 . 
     
     
         9 . The recombinant host cell according to  claim 8 , wherein the host cell is a  Eustigmatophyte  host cell. 
     
     
         10 . The recombinant host cell according to  claim 8 , wherein the host cell is of the genus  Nannochloropsis.    
     
     
         11 . The recombinant host cell according to  claim 8  wherein the nucleic acid sequence is SEQ ID NO: 2 or SEQ ID NO: 3. 
     
     
         12 . The recombinant host cell according to  claim 11  wherein the host cell is a cell of the genus  Nannochloropsis.    
     
     
         13 . The recombinant host cell according to  claim 8 , wherein the heterologous regulatory element is a promoter or terminator. 
     
     
         14 . The recombinant host cell according to  claim 13 , wherein the regulatory element comprises a promoter. 
     
     
         15 . A recombinant host cell comprising a nucleic acid molecule according to  claim 4 . 
     
     
         16 . A nucleic acid molecule according to  claim 1 , wherein said nucleic acid molecule is covalently bound to a detectable label selected from the group consisting of: a fluorescent label, an isotopic label, and an enzymatic label. 
     
     
         17 . The nucleic acid molecule according to  claim 1  bound to a solid support. 
     
     
         18 . The nucleic acid molecule of  claim 17 , wherein the solid support is selected from the group consisting of: a chip, a nucleic acid array, a bead, and a column.