over - expression effect of gene encoding 3-hydroxy-3-methylglutaryl-coa reductase on production of taxol in iranian hazel (corylus avellana l.)

background: sustainable and commercial production of taxol as an anti cancer drug is a critical point to its clinical application. nowadays, hazel because of rapid growth and wide range distribution is considered as an alternative source of taxol. objective: to increase taxol production the cdna encoding 3-hydroxy-3-methylglutaryl-coa reductase (hmgr) from iranian hazel (genebank accession number kf306244, showed by cihmgr) was isolated and over-expressed in pcambia1304 binary vector. the effect of transient over-expression of hmgr in callus and leaf were evaluated on taxol production. methods: the calli was established through the culture of immature cotyledon on murashige and skoog basal medium supplemented with 2, 4-d and ba. the first strand cdna of cihmgr was synthesized by specific primers. enzymatic assay of recombinant cihmgr in e. coli were done by western blott and his-tag affinity techniques. also production of taxol in transformed callus and leaf were evaluated by hplc analysis. results: an open reading frame (orf) with 1698 bp length and a deduced polypeptide with 566 amino acid residues were amplified. the highest and lowest amount of taxol was 0.016 mg/g.dw and 0.004 mg/gdw in transformed calli and untransformed leaves respectively. conclusion: generally the over-expression of hmgr increase the total isoprenoids yield, therefore to have high production of target secondary metabolites (taxol) we need both of network of transformed genes and elicited cell culture.