ISOPENTENYL DIPHOSPHATE ISOMERASE ISOLATED FROM HEVEA BRASILIENSIS AND PRODUCTION METHOD OF RUBBER USING SAME

摘要

PURPOSE: Provided is isopentenyl diphosphate(IPP) isomerase that is isolated from Hevea Brasiliensis. The IPP isomerase catalyzes the conversion between IPP and dimethylallyl diphosphate(DMAPP) each other. Also, a method for manufacturing rubber using the same is provided. CONSTITUTION: A nucleic acid molecule, which encodes Hevea IPP isomerase, is provided. Isopentenyl diphosphate(IPP) isomerase genes are cloned from cDNA library of H. Brasiliensis latex by PCR using two primer(IPP-L1 and IPP-L2) designed based on the Arabidopsis IPP sequence of +606bp to +629bp and an M13 reverse primer. Also, a nucleic acid molecule is separated from Hevea Brasiliensis by hybridizing the nucleic acid molecule under the washing condition of 0.45M sodium chloride, 0.04M sodium citrate and 0.1 % of dudecyl sodium sulfide at a temperature of 45°C. A synthesis method of rubber in vitro comprises the following steps of: a)preparing nucleic acids coding Hevea IPP isomerase; b)producing a transformed host cell by transferring the nucleic acid into a host cell; c)producing IPIHb by culturing the transformed host cell in an optimum medium; and d)synthesizing rubber using the IPIHb in vitro.