Cloning of poly (3-Hydroxybutyrate) synthesis genes from Azotobacter vinelandii into Escherichia coli

The Azotobacter vinelandii is an aerobic, nitrogen-fixing bacterium with the ability to accumulate polyesters such as poly (3-hydroxybutyrate), a particular group of the polyhydroxyalkanoates (pha), when will be cultivated on several carbon sources, including sucrose. Although the capacity of Azotobacter strains to accumulate polyesters is well known, and the genes responsible for their synthesis have recently been identified. However, high production costs have hindered the use of polyhydroxyalkanoates as bioplastics, since their final price is considerably high. The phb gene clusters including phbBAC genes can alternatively be cloned and expressed in the Escherichia coli with the goal to produce high cell density cultivation. We amplified separately 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- TOPO with the subsequent transformation of E. coli strain DH5 alpha.