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Strategy for cloning large gene assemblages as illustrated using the phenylacetate and polyhydroxyalkanoate gene
Belén García1, Elías R Olivera, Angel Sandoval
1Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad de León, Campus de Vegazana s/n, 24007 León, Spain.
Abstract:
We report an easy procedure for isolating chromosome-clustered genes. By following this methodology, the entire set of genes belonging to the phenylacetic acid (PhAc; 18-kb) pathway as well as those required for the synthesis and mobilization of different polyhydroxyalkanoates (PHAs; 6.4 kb) in Pseudomonas putida U were recovered directly from the bacterial chromosome and cloned into a plasmid for the first time. The transformation of different bacteria with these genetic constructions conferred on them the ability to either degrade PhAc or synthesize bioplastics (PHAs).