Porcine γ-synuclein: molecular cloning, expression analysis, chromosomal localization and functional expression

• Published in: Molecular biology reports Vol. 36; no. 5; pp. 971 - 979
• Main Authors: Frandsen, Pernille Munk, Madsen, Lone Bruhn, Bendixen, Christian, Larsen, Knud
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01-05-2009; Springer Netherlands
• Subjects: Amino Acid Sequence; Animal Anatomy; Animal Biochemistry; Animals; Base Sequence; Biomedical and Life Sciences; Cell Line; Chromosomes, Mammalian - metabolism; Cloning, Molecular; DNA, Complementary - genetics; Exons - genetics; gamma-Synuclein - chemistry; gamma-Synuclein - genetics; gamma-Synuclein - metabolism; Gene Expression Regulation; Genome - genetics; Histology; Humans; Intracellular Space - metabolism; Introns - genetics; Life Sciences; Molecular Sequence Data; Morphology; Promoter Regions, Genetic - genetics; Protein Transport; Radiation Hybrid Mapping; Rats; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sequence Alignment; Sus scrofa - genetics; Brain; Animal models; Embryogenesis; γ-synuclein; SNP; Pig
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-008-9270-z

The γ-synuclein protein is involved in breast carcinogenesis and has also been implicated in other forms of cancer and in ocular diseases. Furthermore, γ-synuclein is believed to have a role in certain neurodegenerative diseases, such as Parkinson's disease and Alzheimer's disease. This work reports the cloning and characterization of the porcine (Sus scrofa) γ-synuclein cDNA (SNCG). The SNCG cDNA was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) using oligonucleotide primers derived from in silico sequences. The porcine SNCG cDNA codes for a protein of 126 amino acids which shows a high similarity to bovine (90%), human (87%) and mouse (83%) γ-synuclein. A genomic clone containing the entire porcine SNCG gene was isolated and its genomic organization determined. The gene is composed of five exons, the general structure being observed to be very similar to that of the human SNCG gene. Expression analysis by quantitative real-time RT-PCR revealed the presence of SNCG transcripts in all examined organs and tissues. Differential expression was observed, with very high levels of SNCG mRNA in fat tissue and high expression levels in spleen, cerebellum, frontal cortex and pituitary gland. Expression analysis also showed that porcine SNCG transcripts could be detected in different brain regions during early stages of embryo development. The porcine SNCG orthologue was mapped to chromosome 14q25-q29. The distribution of recombinant porcine γ-synuclein was studied in three different transfected cell lines and the protein was found to be predominantly localized in the cytoplasm.