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Applied and Environmental Microbiology, August 2004, p. 5019-5025, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.5019-5025.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Strategy for Cloning Large Gene Assemblages as Illustrated Using the Phenylacetate and Polyhydroxyalkanoate Gene Clusters

Belén García,1 Elías R. Olivera,1 Ángel Sandoval,1 Elsa Arias-Barrau,1 Sagrario Arias,1 Germán Naharro,2 and José M. Luengo1*

Departamento de Bioquímica y Biología Molecular,1 Departamento de Patología Animal (Sanidad Animal), Facultad de Veterinaria, Universidad de León, 24007 León, Spain2

Received 13 February 2004/ Accepted 21 April 2004

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).

* Corresponding author. Mailing address: Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad de León, Campus de Vegazana s/n, 24007 León, Spain. Phone: 34-987291228. Fax: 34-987291226. E-mail: dbbjlr{at}unileon.es.

Applied and Environmental Microbiology, August 2004, p. 5019-5025, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.5019-5025.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Arias-Barrau, E., Sandoval, A., Naharro, G., Olivera, E. R., Luengo, J. M. (2005). A Two-component Hydroxylase Involved in the Assimilation of 3-Hydroxyphenyl Acetate in Pseudomonas putida. J. Biol. Chem. 280: 26435-26447 [Abstract] [Full Text]  


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