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Molecular cloning, characterization and developmental expression of porcine β-synuclein

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Abstract

The synuclein family includes three known proteins: α-synuclein, β-synuclein and γ-synuclein. β-Synuclein inhibits the aggregation of α-synuclein, a protein involved in Parkinson’s disease. We have cloned and characterized the cDNA sequence for porcine β-synuclein (SNCB) from pig cerebellum using RT-PCR. Expression analysis by quantitative RT-PCR demonstrated that SNCB transcripts were highly abundant in brain tissues. SNCB mRNA was also detected early in embryogenesis and significant increases in transcript levels were observed in several brain tissues during embryo development. Radiation hybrid mapping data indicate that the porcine SNCB maps to the q arm of chromosome 2 (2q21-22). The subcellular localization of recombinant porcine β-synuclein was determined in three different cell types and shown to be cytoplasmic.

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Abbreviations

AD:

Alzheimer’s disease

Bp:

Base pair

CHO:

Chinese hamster ovary

ER:

Endoplasmatic reticulum

EST:

Expressed sequence tag

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

Kb:

Kilobases

Kg:

Kilogram

OLN:

Rat oligodendrocyte cells

ORF:

Open reading frame

PCR:

Polymerase chain reaction

PD:

Parkinson’s disease

pI:

Isoelectric point

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription PCR

SNP:

Single nucleotide polymorphism

U:

Units

UTR:

Untranslated region

Wt:

Wild type

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Acknowledgments

We gratefully acknowledge Dr. Martine Yerle of INRA Toulouse, France for providing the pig-rodent hybrid panel. The authors wish to thank Connie Jakobsen Juhl and Helle Jensen for excellent technical assistance and Dr. Dorothy K. Madsen for critically reading of the manuscript. This study was supported by The Danish Parkinson Association (www.parkinson.dk).

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aarhus, Blichers Alle 20, P.O. Box 50, 8830, Tjele, Denmark

    Knud Larsen, Pernille Munk Frandsen, Lone Bruhn Madsen & Christian Bendixen

Authors
  1. Knud Larsen
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  2. Pernille Munk Frandsen
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  3. Lone Bruhn Madsen
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  4. Christian Bendixen
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Corresponding author

Correspondence to Knud Larsen.

Additional information

The sequence of the porcine SNCB cDNA, encoding the β-synuclein protein, a 5′UTR SNCB cDNA sequence and the entire genomic sequence of SNCB have been submitted to DDBJ/EMBL/GenBank under the accessions numbers EF186678, EU447783, EU621903 and EU552492, respectively.

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Larsen, K., Frandsen, P.M., Madsen, L.B. et al. Molecular cloning, characterization and developmental expression of porcine β-synuclein. Mol Biol Rep 37, 1439–1449 (2010). https://doi.org/10.1007/s11033-009-9531-5

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  • DOI: https://doi.org/10.1007/s11033-009-9531-5

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