Molecular Cloning and Organization of the Coding Region of the Human Norepinephrine Transporter Gene

• Published in: Biochemical and biophysical research communications 1995-10, Vol.215 (3), p.1145-1150
• Main Authors: Porzgen, P., Bonisch, H., Bruss, M.
• Format: Article
• Language: English
• Subjects: Alternative Splicing; Animals; Base Sequence; Carrier Proteins - biosynthesis; Carrier Proteins - chemistry; Carrier Proteins - genetics; Cell Membrane - ultrastructure; Cloning, Molecular; Consensus Sequence; DNA Primers; Dogs; Exons; Genomic Library; Hominidae - genetics; Humans; Introns; Mice; Models, Structural; Molecular Sequence Data; Norepinephrine - metabolism; Norepinephrine Plasma Membrane Transport Proteins; Polymerase Chain Reaction; Protein Structure, Secondary; Rats; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Restriction Mapping; RNA, Messenger - analysis; RNA, Messenger - biosynthesis; Symporters
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1995.2582

A λ phage genomic library was screened with the digoxygenin labeled cDNA of the human norepinephrine transporter (hNET). Six overlapping λ clones were analysed by restriction enzyme analysis and sequencing of the exon-intron boundaries. The coding region of the hNET gene was found to be encoded by 14 exons, spanning 45 kb from the start to the stop codon, disrupted by 13 introns. The organization of the gene is highly homologous to other known neurotransmitter transporter genes. However, the hNET gene differs from the other genes in that it has an additional exon encoding the C-terminus of the protein. The gene structure shows two large introns in the 5′-region and a cluster of 11 exons in the 3′-region. All exon-intron junctions contain the gt/ag consensus splice site. Knowledge of the gene structure of the antidepressant-sensitive hNET should facilitate investigation of its potential role in psychiatric disorders.