Partial cloning and sequencing of the gene encoding the porcine T-cell receptor delta-chain constant region.
The polymerase chain reaction (PCR) was utilized to clone the pig T-cell receptor (TCR) delta-chain constant region-encoding gene (C delta). A cDNA was generated from total RNA preparations of normal pig peripheral blood lymphocytes (PBLs) and a miniature pig peripheral blood cell line (PBLCL 62.G4). The cDNA was used to amplify the porcine TCR C delta gene by PCR using primers chosen by comparing other known C delta sequences for sequence identity. Clones were sequenced and used to determine the primary structure of the porcine TCR C delta chain. A comparison of the nucleotide and deduced amino acid (aa) sequences with the known human, mouse, sheep and cattle sequences revealed that the primary structure of the pig TCR C delta chain has been highly conserved. The immunoglobulin (Ig) domain has two conserved Cys residues and contains a high degree of sequence identity, whereas the hinge region is marked by a high level of diversity. The transmembrane and cytoplasmic regions are also highly conserved, including the presence of the two basic aa, Arg and Lys, in the transmembrane domain. Southern blot analysis has confirmed the presence of one TCR C delta gene in the porcine genome, consistent with similar findings in other species. Thus, the successful cloning and sequencing of the porcine TCR C delta gene should facilitate our understanding of the role of gamma delta T-lymphocytes in the swine immune system.
Department of Molecular Microbiology and Immunology, University of Missouri, School of Medicine, Columbia 65212.
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Grimm
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Misfeldt
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D008969:Molecular Sequence Data; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013552:Swine
D018076:DNA, Complementary; D016692:Receptors, Antigen, T-Cell, gamma-delta