Cis-prenyltransferases from the rubber-producing plants russian dandelion (taraxacum kok-saghyz) and sunflower (helianthus annus)
AssigneePioneer Hi-Bred International, Inc.
InventorsDavid Hallahan, Natalie Keiper-Hrynko
Life of this patent
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Abstract
This invention pertains to nucleic acid fragments encoding plant cis-prenyltransferases. More specifically, this invention pertains to a cis-prenyltransferase homolog from latex of the rubber-producing plant species Taraxacum kok-saghyz (russian dandelion) and a cis-prenyltransferase homolog from the rubber-producing plant species Helianthus annus (sunflower).
Claims25
An isolated nucleic acid molecule encoding a cis-prenyltransferase enzyme, selected from the group consisting of:
- a) an isolated nucleic acid molecule encoding the amino acid sequence as set forth in SEQ ID NOs:4 and 6;
- b) an isolated nucleic acid molecule that hybridizes with (a) under the following hybridization conditions:
- 0.1×SSC, 0.1% SDS, 65° C. and washed with 2×SSC, 0.1% SDS followed by 0.1×SSC, 0.1% SDS;
- or
- an isolated nucleic acid molecule that is complementary to (a) or (b).
An isolated nucleic acid molecule as set forth in SEQ ID NOs:
- 3 and 5.
A polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:6.
An isolated nucleic acid molecule comprising a first nucleotide sequence encoding a polypeptide of at least 301 amino acids that has at least 70% identity based on the Smith-Waterman method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO:4 or a second nucleotide sequence comprising the complement of the first nucleotide sequence, wherein said enzyme has cis-prenyltransferase activity.
An isolated nucleic acid molecule comprising a first nucleotide sequence encoding a polypeptide of at least 168 amino acids that has at least 70% identity based on the Smith-Waterman method of alignment when compared to a polypeptide having the sequence as set forth in SEQ ID NO:6 or a second nucleotide sequence comprising the complement of the first nucleotide sequence, wherein said enzyme has cis-prenyltransferase activity.
A method of obtaining a nucleic acid molecule encoding a cis-prenyltransferase enzyme comprising:
- a) synthesizing at least one oligonucleotide primer corresponding to a portion of the sequence selected from the group consisting of SEQ ID NOs:3 and 5;
- and
- b) amplifying an insert present in a cloning vector using the oligonucleotide primer of step (a);
- wherein the amplified insert encodes a portion of an amino acid sequence encoding a cis-prenyltransferase enzyme.
A method for the production of natural rubber compounds comprising:
- a) providing a transformed host cell comprising:
- (i) suitable levels of isopentenyl pyrophate;
- and
- (ii) a cis-prenyltransferase gene selected from the group consisting of SEQ ID NOs:
- 3 and 5, wherein said genes are operably linked to suitable regulatory sequences;
- and
- b) growing the transformed host cell of (a) under conditions whereby a natural rubber compound is produced.
A method for the identification of a polypeptide having cis-prenyltransferase activity in a rubber-producing plant comprising:
- (a) obtaining the amino acid sequence of a polypeptide suspected of having cis-prenyltransferase activity;
- and
- (b) aligning the amino acid sequence of step (a) with the amino acid sequence of a cis-prenyltransferase consensus sequence selected from the group consisting of SEQ ID NO:4, 6, 8, 9, and 10, wherein the alignment shows the presence of conserved domains I, IV, and V (SEQ ID NOs:
- 38-40).
A method for the identification of a polypeptide having cis-prenyltransferase activity in a rubber-producing plant comprising:
- (a) obtaining the amino acid sequence of a polypeptide suspected of having cis-prenyltransferase activity;
- and
- (b) aligning the amino acid sequence of step (a) with the amino acid sequence of a cis-prenyltransferase consensus sequence selected from the group consisting of SEQ ID NO:4, 6, 8, 9, and 10, wherein the alignment shows a sequence of at least about 50 non-conserved amino acids present between the absolutely conserved tyrosine of Domain IV and the first of the absolutely conserved arginine residue of Domain V.
Dependencies are inferred from the claim text.