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Cloning and characterization of the porcine DBC1 gene encoding deleted in bladder cancer

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Abstract

Deleted in bladder cancer 1 (DBC1) is a tumour suppressor which is involved in the regulation of cell growth and programmed cell death. In this study we report the cloning and characterization of porcine DBC1 cDNA. RT-PCR cloning produced a cDNA with an open reading frame of 2,283 bp encoding a polypeptide of 761 amino acids with a predicted molecular mass of 88.6 kDa and estimated isoelectric point of 9.1. The encoded pig DBC1 protein shows a very high amino acid similarity to human (99 %) and to mouse (98 %) DBC1. The porcine DBC1 gene was mapped to chromosome 1. The nucleotide sequence of the promoter displayed a high degree of conservation of elements responsible for neuron-specific expression. The porcine DBC1 gene was found to be highly expressed in brain tissues. The methylation status of the porcine DBC1 gene was examined in brain and liver by bisulfite sequencing. Methylation percentages of 53–61 were observed for the gene body whereas significantly lower values (1–4 %) were found in exon 1 and the promoter sequence of DBC1. The sequences of the porcine DBC1 cDNA and the DBC1 promoter and exon 1 sequence have been submitted to DDBJ/EMBL/GenBank under the accession numbers KF733442 and KJ396193, respectively.

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Acknowledgments

The authors wish to thank Connie Jakobsen Juhl and Bente Flügel for excellent technical assistance and Ms. Janne Hansen for critical reading of the manuscript. The work was supported financially by a Grant from the Danish Agency for Science, Technology and Innovation (274-09-0299).

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Science and Technology, Aarhus University, Blichers Alle 20, P.O. Box 50, 8830, Tjele, Denmark

    Knud Larsen, Jamal Momeni, Leila Farajzadeh & Christian Bendixen

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  1. Knud Larsen
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  2. Jamal Momeni
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  3. Leila Farajzadeh
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  4. Christian Bendixen
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Correspondence to Knud Larsen.

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Larsen, K., Momeni, J., Farajzadeh, L. et al. Cloning and characterization of the porcine DBC1 gene encoding deleted in bladder cancer. Mol Biol Rep 42, 383–391 (2015). https://doi.org/10.1007/s11033-014-3779-0

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  • DOI: https://doi.org/10.1007/s11033-014-3779-0

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