Cloning of poly (3-Hydroxybutyrate) synthesis genes from Azotobacter

The Azotobacter vinelandii is an aerobic, nitrogen-fixing bacterium with the ability toaccumulate polyesters such as poly (3-hydroxybutyrate), a particular group of thepolyhydroxyalkanoates (pha), when will be cultivated on several carbon sources,including sucrose. Although the capacity of Azotobacter strains to accumulate polyestersis well known, and the genes responsible for their synthesis have recently beenidentified. However, high production costs have hindered the use ofpolyhydroxyalkanoates as bioplastics, since their final price is considerably high. Thephb gene clusters including phbBAC genes can alternatively be cloned and expressed inthe Escherichia coli with the goal to produce high cell density cultivation. We amplifiedseparately the phbB, phbA and phbC genes from extracted Azotobacter genome with the designed primers and could clone the fragments in the cloning vector pCR 2.1- TOPOwith the subsequent transformation of E. coli strain DH5.