Lcb1 subunit of serine palmitoyltransferase
AssigneeE. I. Du Pont de Nemours and Company
InventorsRebecca E. Cahoon, Anthony J. Kinney, J. Antoni Rafalski
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Abstract
This invention relates to an isolated nucleic acid fragment encoding a serine palmitoyltransferase Lcb1 subunit. The invention also relates to the construction of a chimeric gene encoding all or a portion of the serine palmitoyltransferase Lcb1 subunit, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the serine palmitoyltransferase Lcb1 subunit in a transformed host cell.
Claims11
An isolated nucleic acid fragment encoding all or a substantial portion of a serine palmitoyltransferase Lcb1 subunit comprising a member selected from the group consisting of:
- (a) an isolated nucleic acid fragment encoding all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10 and SEQ ID NO:
- 12;
- (b) an isolated nucleic acid fragment that is substantially similar to an isolated nucleic acid fragment encoding all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10 and SEQ ID NO:
- 12;
- and
- (c) an isolated nucleic acid fragment that is complementary to (a) or (b).
A serine palmitoyltransferase Lcb1 subunit polypeptide comprising all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, SEQ ID NO:
- 10 and SEQ ID NO:
- 12.
A method of obtaining a nucleic acid fragment encoding a substantial portion of an amino acid sequence encoding a serine palmitoyltransferase Lcb1 subunit comprising:
- (a) synthesizing an oligonucleotide primer corresponding to a portion of the sequence set forth in any of SEQ ID NOs:
- 1, 3, 5, 7, 9 and 11;
- and
- (b) amplifying a cDNA insert present in a cloning vector using the oligonucleotide primer of step (a) and a primer representing sequences of the cloning vector
- wherein the amplified nucleic acid fragment encodes a substantial portion of an amino acid sequence encoding a serine palmitoyltransferase Lcb1 subunit.
A method for evaluating at least one compound for its ability to inhibit the activity of a serine palmitoyltransferase Lcb1 subunit, the method comprising the steps of:
- (a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding a serine palmitoyltransferase Lcb1 subunit, operably linked to suitable regulatory sequences;
- (b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of the serine palmitoyltransferase Lcb1 subunit encoded by the operably linked nucleic acid fragment in the transformed host cell;
- (c) optionally purifying the serine palmitoyltransferase Lcb1 subunit expressed by the transformed host cell;
- (d) treating the serine palmitoyltransferase Lcb1 subunit with a compound to be tested;
- and
- (e) comparing the activity of the serine palmitoyltransferase Lcb1 subunit that has been treated with a test compound to the activity of an untreated serine palmitoyltransferase Lcb1 subunit,
- thereby selecting compounds with potential for inhibitory activity.
Dependencies are inferred from the claim text.