Document Type : Brief Report
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
University of Tabriz, Faculty of Agriculture, Department of Soil Science, P.O. Box 5166616471, Tabriz, I.R. Iran.
Federal Research Institute of Nutrition and Food, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.
Phosphatase (APase) enzymes including phytases have broad applications in diagnostic kits, poultry
feeds, biofertilizers and plant nutrition. Because of high levels of sequence diversity among phosphatases,
an efficient functional screening method is a crucial requirement for the isolation of the encoding
genes. This study reports a functional cloning screening method for the isolation of APase-encoding genes
from bacterial genomic libraries in a medium containing a chromogenic substrate. The method was optimized
to distinguish the desired signal from the background chromosomal APase activity. This screening method led to the isolation of two novel APase-encoding genes from Pseudomonas putida with no similarities to the known genes in the databases, indicating successful implementation of the developed method.