Genomic structure and mapping of the chromosomal gene for transcobalamin I (TCN1): comparison to human intrinsic factor.
Transcobalamin I (TCI) is a vitamin B12 binding protein that is found in the secondary granules of mature neutrophils. The expression of the gene for TCI (TCN1) within neutrophils has been shown to be restricted to the later stages of myeloid development and can therefore be used as a marker for granulocyte differentiation. To study transcriptional control regions important in late stage myeloid gene regulation the genomic sequence for TCN1 has been cloned. Clones were isolated from a genomic library constructed in Charon 4A using homologous full-length cDNA probes. Southern blot analysis showed the gene to reside on five EcoRI fragments totaling 14 kb in length. Two overlapping phage clones, containing the entire 14 kb, were isolated and the introns and exons were mapped using Southern blotting and dideoxy sequencing of subclones. The cDNA is represented by nine exons contained within 12 kb of genomic DNA. Comparison of the genomic structure to gastric intrinsic factor (GIF), another vitamin B12 binding protein, revealed a strikingly similar intron/exon structure, with several positionally conserved splice sites. The gene was localized to chromosome 11 using in situ hybridization.
Johnston
J
J
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Yang-Feng
T
T
Berliner
N
N
D000595:Amino Acid Sequence; D015139:Blotting, Southern; D002871:Chromosome Banding; D002874:Chromosome Mapping; D002880:Chromosomes, Human, Pair 11; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D006801:Humans; D007437:Intrinsic Factor / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014155:Transcobalamins / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D014155:Transcobalamins; D004247:DNA; D007437:Intrinsic Factor