Patent
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.
1. An isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, and a full length complement of any thereof; further wherein the nucleic acid sequence is operably linked to a heterologous regulatory element.
2. The isolated 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 isolated nucleic acid molecule according to claim 1 , wherein the heterologous regulatory element is a promoter or terminator.
4. The isolated nucleic acid molecule according to claim 3 , wherein the regulatory element comprises a promoter.
5. The isolated nucleic acid molecule according to claim 1 , wherein the nucleic acid sequence further comprises a coding sequence for a polypeptide.
6. The isolated nucleic acid molecule according to claim 5 , wherein the coding sequence for a polypeptide is not from the same species as the isolated nucleic acid sequence.
7. The isolated nucleic acid molecule according to claim 6 , wherein the coding sequence for a polypeptide is not from a Nannochloropsis species.
8. A recombinant host cell comprising the nucleic acid molecule according to claim 1 .
9. The recombinant host cell according to claim 8 , wherein the host cell is a heterokont.
10. The recombinant host cell according to claim 9 , wherein the host cell is a Nannochloropsis cell.
11. An isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, and a full length complement of any thereof; wherein said nucleic acid molecule is less than 250 nucleotides in length and is covalently bound to a detectable label selected from the group consisting of: a fluorescent label, an isotopic label, and an enzymatic label.
12. The isolated nucleic acid molecule according to claim 11 , wherein said isolated nucleic acid molecule is bound to a solid support.
13. The isolated nucleic acid molecule of claim 12 wherein the solid support is selected from the group consisting of: a chip, a nucleic acid array, a bead, and a column.
14. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid sequence is SEQ ID NO:3.
15. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid sequence is SEQ ID NO:5.