Cloning of the beta 2-microglobulin gene in the zebrafish.
The beta 2-microglobulin (beta 2m) is a protein found in the serum in a free form and on the cell surface in a form noncovalently associated with the alpha chain of the class I major histocompatibility complex (Mhc) molecules. In mammals, the beta 2m-encoding gene (B2m) is found on a chromosome different from the Mhc proper. We have isolated and characterized the B2m gene of the zebrafish, Brachydanio rerio, family Cyprinidae. We obtained both cDNA and genomic clones of the Brre-B2m gene. The cDNA clones contained the entire coding sequence, the entire 3' untranslated (UT) region, and at least part of the 5'UT region. The genomic clone contained the entire Brre-B2m gene. The coding sequence specifies 97 amino acid residues of the mature protein so that the zebrafish beta 2m is two residues shorter than human and one residue shorter than cattle, fowl, or turkey beta 2m (codons at positions 85 and 86 have been deleted in the Brre-B2m gene). The amino acid and nucleotide sequence similarities between zebrafish and human beta 2m (B2m) are 45% and 59%, respectively. Approximately 24% of the positions are invariant and an additional 9% show only conservative substitutions in comparisons which include all known beta 2m sequences (fish, avian, and mammalian). Most of the conserved positions are in the beta strands (some 47% of the beta-strand positions are conserved in the three vertebrate classes). The Brre-B2m gene consists of four exons separated by three introns. All of the introns are considerably shorter than the corresponding introns in the mammalian B2m genes. The coding sequences of the cDNA and the genomic clones are almost identical but the sequences of the 3'UT regions differ at 1.7% of the sites, suggesting that the genes borne by these clones might have diverged at least 0.7 million years (my) ago. In contrast to the human B2m gene, the Brre-B2m gene shows no bias in the distribution of the CpG dinucleotides: the dinucleotides are distributed evenly along the entire available sequence. The haploid genome of the zebrafish contains only one copy of the B2m gene.
Ono
H
H
Max-Planck-Institut für Biologie, Abteilung Immungenetik, Tübingen, Germany.
Figueroa
F
F
O'hUigin
C
C
Klein
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D015183:Restriction Mapping; D016415:Sequence Alignment; D012995:Solubility; D015027:Zebrafish / Q000235:genetics* / Q000276:immunology; D001613:beta 2-Microglobulin / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D001613:beta 2-Microglobulin; D004247:DNA