Method for producing trans-polyisoprenoid, vector, transgenic plant, method for producing pneumatic tire and method for producing rubber product
The present invention aims to provide a method for producing a trans-polyisoprenoid which can increase trans rubber production. The present invention is directed to a method for producing a trans-polyisoprenoid in vitro, which involves the use of a gene coding for a trans-prenyltransferase (tPT) family protein and further involves the use of rubber particles bound to a protein encoded by the gene, or a method for producing a trans-polyisoprenoid, which includes introducing into a plant a vector including a promotor having a promoter activity that drives laticifer-specific gene expression and a gene coding for a tPT family protein linked to the promotor to express a protein encoded by the gene specifically in laticifers.
1. A method for producing a trans-polyisoprenoid, the method comprising:
binding a protein expressed from a gene coding for a trans-prenyltransferase (tPT) family protein to rubber particles in vitro,
wherein said binding is that said protein is fully or partially incorporated into the rubber particle, or the protein is inserted into a membrane structure of the rubber particle, and
wherein the binding comprises performing protein synthesis in the presence of both rubber particles and a cell-free protein synthesis solution containing an mRNA coding for a trans-prenyltransferase (tPT) family protein to bind the tPT family protein to the rubber particles.
2. The method for producing a trans-polyisoprenoid according to claim 1 ,
wherein the gene coding for a trans-prenyltransferase (tPT) family protein is derived from a plant.
3. The method for producing a trans-polyisoprenoid according to claim 2 ,
wherein the gene coding for a trans-prenyltransferase (tPT) family protein is derived from a rubber-producing plant.
4. The method for producing a trans-polyisoprenoid according to claim 3 ,
wherein the gene coding for a trans-prenyltransferase (tPT) family protein is derived from Hevea brasiliensis.
5. The method for producing a trans-polyisoprenoid according to claim 1 ,
wherein the cell-free protein synthesis solution comprises a germ extract.
6. The method for producing a trans-polyisoprenoid according to claim 5 ,
wherein the germ extract is derived from wheat.
7. The method for producing a trans-polyisoprenoid according to claim 1 ,
wherein the rubber particles are present in the cell-free protein synthesis solution at a concentration of 5 to 50 g/L.