ISOPENTENYL DIPHOSPHATE ISOMERASE FROM HEVEA BRASILIENSIS AND RUBBER PRODUCING METHOD USING THE SAME

摘要

From the cDNA library of the latex of the Brazilian rubber tree (Hevea brasiliensis) it was cloned a cDNA clone iso-pentenyl encoded phosphate (IPP) isomerase (EC 5.3.3.2). The clone has a contiguous open reading frame encoded by of the expected molecular weight 26.7 kDa 234 amino acid peptide. The deduced protein is acidic with an isoelectric point of 4.7 and shows another IPP isomerase with high sequence identity. The recombinant protein expressed in E. coli (Escherichia coli) represents the IPP isomerase activity. Allyl initiator to the use of rubber particles (WRP) washing to remove phosphoric acid was added to IPP isomerase in the reaction mixture, which was run the synthesis in vitro analysis of the rubber. Recombinant IPP isomerase is shown the result that the catalytic effect of promoting the conversion to DMAPP in the basic five-carbon isoprene units important activation step of the IPP in rubber biosynthesis. The IPP isomerase by Southern analysis (Southern analysis) confirmed that encoded by the two genes of the Brazilian rubber tree. Northern blot analysis (Northern blot analysis) single hybridization was detected two transcripts (1.2 kb and 0.6 kb) of the same size from leaf tissues, while easing the band is detected from the fluid in the. The analysis of the RNA extracted from leaf tissue with lotion bursting results wounds of trees embellish wounds four days (nail) showed that it does not alter the transcription level of IPP isomerase.