isolation and cloning of sat gene in papaver somniferumand evaluation of its transient expression

abstract benzylisoquinoline alkaloids are a diverse group of nitrogenous compounds which is found in about 20% of plant species. isolation of effective genes involved in morphine biosynthesis of opium poppy is very important in the production of specific which can be achieved using metabolic engineering techniques. in this biosynthesis pathway, the key enzyme sat is involved in the conversion of salutaridinol 7-o-acetyltransferase (ec 2.3.1.150) to salutaridinol-7-o-acetate, which is the immediate precursor of thebaine. in this project, the gene encoding this enzyme was isolated using primers which were designed on the basis of the gene sequence available on data banks (ncbi) for papaver somniferum. this gene is then cloned in expression vectors under the control of camv 355 promoter. the result of this cloning was confirmed using different molecular methods such as enzyme digestion and pcr. agro infiltration method was also used for transient expression of sat gene. the result of evaluation showed that morphine and codeine were only produced in the leaves of transgenic plants containing sat transgen.