Cloning two PepT1 cDNA fragments of common carp, Cyprinus carpio (Actinopterygii: Cypriniformes: Cyprinidae)
2009
Ostaszewska, T.,Warsaw University of Life Sciences (Poland). Dept. of Ichthyobiology and Fisheries | Szatkowska, I.,West Pomeranian University of Technology, Szczecin (Poland). Dept of Molecular Cytogenetics | Verri, T.,University of Salento, Lecce (Italy). Lab. of General Physiology | Dabrowski, K.,Ohio State University, Columbus (USA). School of Environ. and Natural Res. | Romano, A. | Barca, A.,University of Salento, Lecce (Italy). Lab. of General Physiology | Muszynska, M. | Dybus, A.,West Pomeranian University of Technology, Szczecin (Poland). Dept of Molecular Cytogenetics | Grochowski, P. | Kamaszewski, M.,Warsaw University of Life Sciences (Poland). Dept. of Ichthyobiology and Fisheries
Oligopeptides are a new and promising alternative source of amino acids in animal as well as in human nutrition. The aim of this paper was to sequence the PepT1 (SLC15A1) in common carp, Cyprinus carpio. Intestine samples were isolated from six-week old common carp. Total RNA was isolated using a Trizol method. Reverse transcription was used to synthesize cDNA. Two different pairs of primers were designed, according to the zebrafish (Danio rerio) PepT1 sequence, and used for PCR. The amplified DNA was isolated by electrophoresis, cloned (pCRII-TOPO vectors), sequenced, and subjected to in silico analysis. Two nucleotide fragments of the PepT1 gene were obtained and analyzed using bioinformatic tools. Both fragments showed a high degree of homology with the known PepT1 genes of other animals, and were deposited in GenBank (FJ556590, FJ529670). The sequenced fragments of the common carp PepT1 gene will allow evaluation of PepT1 expression in the intestines of fish fed diets containing various forms of protein
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