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1.
IJB-Iranian Journal of Biotechnology. 2010; 8 (4): 275-279
Dans Anglais | IMEMR | ID: emr-145280

Résumé

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


Sujets)
Phytase/génétique , Gènes , Séquence nucléotidique , Pseudomonas putida
2.
Saudi Medical Journal. 2006; 27 (9): 1397-1400
Dans Anglais | IMEMR | ID: emr-80938

Résumé

To develop a molecular technique that is fast and reliable in detecting porcine contamination or ingredients in foods. The method applied involved DNA amplification using polymerase chain reaction [PCR] technology. Thus, the sequence of a certain gene found uniquely in pork was identified and its sequence was used to design specific primers for the PCR. The extraction of DNA was optimized in respect to PCR and detection limits were established. The optimized method was then used to identify pork in food products obtained from various local hypermarkets. The latest results were confirmed in triplicates on the 20th April 2006 at the Molecular Biology Laboratory, International Islamic University, Malaysia. The method was shown to be robust and reliable. Out of 30 food samples not expected to contain pork material, 3 samples were shown to be contaminated with pork material; 2 chocolates and one chicken nugget. We observed that 2 food products that were labeled as halal showed positive for porcine ingredients, while another that did not have any halal logo but originated from outside Malaysia and exported to many Middle Eastern nations also showed positive


Sujets)
Réaction de polymérisation en chaîne/médecine vétérinaire , Réaction de polymérisation en chaîne/méthodes , Manipulation des aliments , Fragmentation de l'ADN , Données de séquences moléculaires , Sensibilité et spécificité
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