Molecular cloning and characterization of the bovine and human tuftelin genes

The bovine tuftelin gene was cloned and its structure determined by DNA seq uence analysis and comparison to bovine tuftelin cDNA. The analyses demonst rated that the cDNA contains a 1014 bp open reading frame encoding a protei n of 338 residues with a calculated molecular weight of 38,630 kDa and an i soelectric point of 5.85. Although similar, these results differ from those previously published [Deutsch et al. (1991) J. Biol. Chem. 266, 16021-1602 8] which contained a different conceptual amino acid sequence for the carbo xy terminal region and identification of a different termination codon beca use of the absence of a single guanine residue in the published sequence, T he protein does not appear to share homology or domain motifs with any ethe r known protein, The bovine gene consists of 13 exons ranging in size from 66 to 1531 bp, the latter containing the encoded carboxy terminal and 3' un translated regions, These exons are embedded in greater than 28 kbp of geno mic DNA and codons are generally not divided at exon/intron borders, Sequen ce analysis of the cDNA and products produced by reverse transcriptase/poly merase chain reaction demonstrated that exons 2, 5 and 6 are alternatively spliced. The 3' portion of the human gene was also isolated and characteriz ed by DNA sequencing, which demonstrated agreement between the bovine and h uman sequences in the segment in question, The difference between the prese ntly reported sequence and that of the previously published one suggests th e possibility of an unusual type of polymorphism which would result in mark edly different amino acid sequences at the carboxy terminal region of the p rotein. The human tuftelin gene was localized to chromosome 1q21 by in situ hybridization.