جداسازی و کلونینگ ژن ایزوفلاون سنتتاز از گیاه سویا رقم ویلیامز III

Isoflavonoids are a group of flavonoid compound which produced from flavanon precursors in legums, by isoflavone synthase (IFS) enzyme. The most important isoflavonoids finding in soybean are genistein and daidzein that have many medicinal impacts. Investigations have revealed that regularly using these compounds lowered the risk to come down with hormone-related cancers, besides menopausal symptoms and reduction in osteoprosis, balance in cholesterol bloods level, improving in heart-coronary diseases are benefits that are related to using these compounds. In this investigation in order to isolate isoflavone synthase, at first total RNA was extracted and cDNA was synthesized, subsequently. IFS gene was isolated and subjected to PCR by specific primers which have the HindIII and XbaI at their 5′ ends. Subsequently, amplified gene after extracting on agarose gel was cloned in restriction site of XbaI/HnidIII of pET28a+. At the last step, recombinant plasmids were approved by PCR and enzymatic digestion. Finally, the mentioned vector can be used either as an expression vector in prokaryotic hosts or introduced into plants by agrobacterium or gene gun-mediated transformations.