Isolation and Cloning of Three Genes of Polyhydroxybutirate Operon Producing Biodegedable Plastic

Polyhydroxybutyrate (PHB) is a polyhydroxy alkanote (PHA) which produces in ralstonia eutropha by three enzymes, including Beta-ketothiolase, NADPH-dependent acetoacetyl-CoA reductase and PHB synthase. These three enzymes are encoded by phbA, phbB and phbC respectively. There is considerable interest in Polyhidroxybutirate, since it can be used as biodegradable plastic, which under optimal conditions are degraded completely to CO2 and H2O. In this research phb operon was isolated from ralstonia genome using polymerase chain reaction. Specific primers were designed for PHB isolation. The PCR product was purified and cloned. Accuracy of isolation and cloning of three PHB genes were verified by nested PCR and digestion reaction. It is expected that cloning of PHB genes in expression and transformation vectors can be caused to cost effective production of Polyhydroxybutyrate as a bio-degradable plastic in plants.