Cyclopropane-fatty-acyl-phospholipid synthase

Abstract

This invention relates to an isolated nucleic acid fragment encoding a cyclopropane synthetase. The invention also relates to the construction of a chimeric gene encoding all or a portion of the cyclopropane synthetase, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the cyclopropane synthetase in a transformed host cell.

What is claimed is:  
     
         1 . An isolated nucleic acid fragment encoding all or a substantial portion of a cyclopropane synthetase 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 SEQ ID NO:2;    (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 SEQ ID NO:2; and    (c) an isolated nucleic acid fragment that is complementary to (a) or (b).    
     
     
         2 . The isolated nucleic acid fragment of  claim 1  wherein the nucleic acid fragment encodes an amino acid sequence that is greater than 80% similar to the amino acid sequence set forth in SEQ ID NO:2.  
     
     
         3 . The isolated nucleic acid fragment of  claim 1  wherein the nucleic acid fragment is complementary to a nucleic acid fragment that encodes an amino acid sequence that is greater than 80% similar to the amino acid sequences set forth in SEQ ID NO:2.  
     
     
         4 . The isolated nucleic acid fragment of  claim 1  wherein the nucleic acid fragment hybridizes under stringent conditions to a nucleic acid fragment that encodes the amino acid sequence set forth in SEQ ID NO:2.  
     
     
         5 . The isolated nucleic acid fragment of  claim 4  wherein the nucleic acid fragment encodes a functional RNA.  
     
     
         6 . The isolated nucleic acid fragment of  claim 1  wherein the nucleotide sequence of the fragment comprises all or a substantial portion of the sequence set forth in SEQ ID NO: 1.  
     
     
         7 . A chimeric gene comprising the nucleic acid fragment of  claim 1  operably linked to suitable regulatory sequences.  
     
     
         8 . A transformed host cell comprising the chimeric gene of  claim 7 .  
     
     
         9 . A cyclopropane synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:2.  
     
     
         10 . An isolated nucleic acid fragment encoding all or a substantial portion of a cyclopropane synthetase 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 SEQ ID NO:4;    (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 SEQ ID NO:4; and    (c) an isolated nucleic acid fragment that is complementary to (a) or (b).    
     
     
         11 . The isolated nucleic acid fragment of  claim 10  wherein the nucleic acid fragment encodes an amino acid sequence that is greater than 80% similar to the amino acid sequence set forth in SEQ ID NO:4.  
     
     
         12 . The isolated nucleic acid fragment of  claim 10  wherein the nucleic acid fragment is complementary to a nucleic acid fragment that encodes an amino acid sequence that is greater than 80% similar to the amino acid sequences set forth in SEQ ID NO:4.  
     
     
         13 . The isolated nucleic acid fragment of  claim 10  wherein the nucleic acid fragment hybridizes under stringent conditions to a nucleic acid fragment that encodes the amino acid sequence set forth in SEQ ID NO:4.  
     
     
         14 . The isolated nucleic acid fragment of  claim 13  wherein the nucleic acid fragment encodes a functional RNA.  
     
     
         15 . The isolated nucleic acid fragment of  claim 10  wherein the nucleotide sequence of the fragment comprises all or a substantial portion of the sequence set forth in SEQ ID NO:3.  
     
     
         16 . A chimeric gene comprising the nucleic acid fragment of  claim 10  operably linked to suitable regulatory sequences.  
     
     
         17 . A transformed host cell comprising the chimeric gene of  claim 16 .  
     
     
         18 . A cyclopropane synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:4.  
     
     
         19 . An isolated nucleic acid fragment encoding all or a substantial portion of a cyclopropane synthetase 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 SEQ ID NO:6;    (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 SEQ ID NO:6; and    (c) an isolated nucleic acid fragment that is complementary to (a) or (b).    
     
     
         20 . The isolated nucleic acid fragment of  claim 1  wherein the nucleic acid fragment encodes an amino acid sequence that is greater than 80% similar to the amino acid sequence set forth in SEQ ID NO:6.  
     
     
         21 . The isolated nucleic acid fragment of  claim 19  wherein the nucleic acid fragment is complementary to a nucleic acid fragment that encodes an amino acid sequence that is greater than 80% similar to the amino acid sequences set forth in SEQ ID NO:6.  
     
     
         22 . The isolated nucleic acid fragment of  claim 19  wherein the nucleic acid fragment hybridizes under stringent conditions to a nucleic acid fragment that encodes the amino acid sequence set forth in SEQ ID NO:6.  
     
     
         23 . The isolated nucleic acid fragment of  claim 22  wherein the nucleic acid fragment encodes a functional RNA.  
     
     
         24 . The isolated nucleic acid fragment of  claim 19  wherein the nucleotide sequence of the fragment comprises all or a substantial portion of the sequence set forth in SEQ ID NO:5.  
     
     
         25 . A chimeric gene comprising the nucleic acid fragment of  claim 19  operably linked to suitable regulatory sequences.  
     
     
         26 . A transformed host cell comprising the chimeric gene of  claim 25 .  
     
     
         27 . A cyclopropane synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:6.  
     
     
         28 . An isolated nucleic acid fragment encoding all or a substantial portion of a cyclopropane synthetase 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 SEQ ID NO:8;    (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 SEQ ID NO:8; and    (c) an isolated nucleic acid fragment that is complementary to (a) or (b).    
     
     
         29 . The isolated nucleic acid fragment of  claim 28  wherein the nucleic acid fragment encodes an amino acid sequence that is greater than 80% similar to the amino acid sequence set forth in SEQ ID NO:8.  
     
     
         30 . The isolated nucleic acid fragment of  claim 28  wherein the nucleic acid fragment is complementary to a nucleic acid fragment that encodes an amino acid sequence that is greater than 80% similar to the amino acid sequences set forth in SEQ ID NO:8.  
     
     
         31 . The isolated nucleic acid fragment of  claim 28  wherein the nucleic acid fragment hybridizes under stringent conditions to a nucleic acid fragment that encodes the amino acid sequence set forth in SEQ ID NO:8.  
     
     
         32 . The isolated nucleic acid fragment of  claim 31  wherein the nucleic acid fragment encodes a functional RNA.  
     
     
         33 . The isolated nucleic acid fragment of  claim 31  wherein the nucleotide sequence of the fragment comprises all or a substantial portion of the sequence set forth in SEQ ID NO:7.  
     
     
         34 . A chimeric gene comprising the nucleic acid fragment of  claim 28  operably linked to suitable regulatory sequences.  
     
     
         35 . A transformed host cell comprising the chimeric gene of  claim 34 .  
     
     
         36 . A cyclopropane synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:8.  
     
     
         37 . An isolated nucleic acid fragment encoding all or a substantial portion of a cyclopropane synthetase 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 SEQ ID NO:10;    (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 SEQ ID NO:10; and    (c) an isolated nucleic acid fragment that is complementary to (a) or (b).    
     
     
         38 . The isolated nucleic acid fragment of  claim 37  wherein the nucleic acid fragment encodes an amino acid sequence that is greater than 80% similar to the amino acid sequence set forth in SEQ ID NO: 10.  
     
     
         39 . The isolated nucleic acid fragment of  claim 37  wherein the nucleic acid fragment is complementary to a nucleic acid fragment that encodes an amino acid sequence that is greater than 80% similar to the amino acid sequences set forth in SEQ ID NO:10.  
     
     
         40 . The isolated nucleic acid fragment of  claim 37  wherein the nucleic acid fragment hybridizes under stringent conditions to a nucleic acid fragment that encodes the amino acid sequence set forth in SEQ ID NO:10.  
     
     
         41 . The isolated nucleic acid fragment of  claim 40  wherein the nucleic acid fragment encodes a functional RNA.  
     
     
         42 . The isolated nucleic acid fragment of  claim 37  wherein the nucleotide sequence of the fragment comprises all or a substantial portion of the sequence set forth in SEQ ID NO:9.  
     
     
         43 . A chimeric gene comprising the nucleic acid fragment of  claim 37  operably linked to suitable regulatory sequences.  
     
     
         44 . A transformed host cell comprising the chimeric gene of  claim 43 .  
     
     
         45 . A cyclopropane synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:10.  
     
     
         46 . A method of altering the level of expression of a cyclopropane synthetase in a host cell comprising: 
 (a) transforming a host cell with the chimeric gene of any of claims  3 ,  12 ,  21 ,  30  and  39 ; and    (b) growing the transformed host cell produced in step (a) under conditions that are suitable for expression of the chimeric gene    wherein expression of the chimeric gene results in production of altered levels of a cyclopropane synthetase in the transformed host cell.    
     
     
         47 . A method of obtaining a nucleic acid fragment encoding all or a substantial portion of the amino acid sequence encoding a cyclopropane synthetase comprising: 
 (a) probing a cDNA or genomic library with the nucleic acid fragment of any of claims  1 ,  10 ,  19 ,  28  and  37 ;    (b) identifying a DNA clone that hybridizes with the nucleic acid fragment of any of claims  1 ,  10 ,  19 ,  28  and  37 ;    (c) isolating the DNA clone identified in step (b); and    (d) sequencing the cDNA or genomic fragment that comprises the clone isolated in step (c)    wherein the sequenced nucleic acid fragment encodes all or a substantial portion of the amino acid sequence encoding a cyclopropane synthetase.    
     
     
         48 . A method of obtaining a nucleic acid fragment encoding a substantial portion of an amino acid sequence encoding a cyclopropane synthetase 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 and 9; 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 cyclopropane synthetase.    
     
     
         49 . The product of the method of  claim 47 .  
     
     
         50 . The product of the method of  claim 48.