Strategy for cloning large gene assemblages as illustrated using the phenylacetate and polyhydroxyalkanoate gene clusters.
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).
Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad de León, Campus de Vegazana s/n, 24007 León, Spain.
Belén
B
García
Elías R
ER
Olivera
Angel
A
Sandoval
Elsa
E
Arias-Barrau
Sagrario
S
Arias
Germán
G
Naharro
José M
JM
Luengo
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D005810:Multigene Family / Q000235:genetics*; D010648:Phenylacetates / Q000378:metabolism*; D010957:Plasmids; D011091:Polyesters / Q000378:metabolism*; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D010648:Phenylacetates; D011091:Polyesters; C025136:phenylacetic acid