Journal article
Cloning, sequencing and expression of the bovine CD3ε and TCR-ζ chains, two invariant components of the T-cell receptor complex
Gene, Vol.169(2), pp.165-171
1996
Handle:
https://hdl.handle.net/2376/114506
PMID: 8647441
Abstract
CD3ε and the ζ-chain of the bovine T-cell receptor (TCR) are two invariant molecules with an important role in signal transduction via the TCR/CD3 complex. The nucleotide sequence of a bovine
CD3ε cDNA clone containing the complete coding sequence was determined and the deduced amino acid (aa) sequence compared to that of other species. The cytoplasmic domains of the different CD3ε clearly show a higher degree of conservation than the extracellular domains. Bovine CD3ε produced in
Escherichia coli using different bacterial expression vectors was recognised by antibodies (Ab) directed against the intracytoplasmic domain of human CD3ε. A partial bovine
TCRζ-chain cDNA was generated by the polymerase chain reaction (PCR) using primers that were based on sequences that are conserved between different species; 3′ and 5′ RACE-PCR were carried out to obtain the complete
TCRζ-chain cDNA sequence. A comparison of the predicted TCRζ-chain aa sequence reveals that the GDP/GTP-binding motif, which is conserved in other species, shows marked differences in the bovine and ovine TCRζ-chains. In contrast to CD3ε, the short extracellular domain of the TCRζ-chain is 100% conserved between the different species and the transmembrane domain also shows a high degree of identity. Ab were raised against the TCRζ-chain, produced as a glutathione S-transferase fusion protein in
E. coli, and were used in Western blot analysis to further characterise
TCRζ-chain expression in T-cells. These reagents provide valuable tools for the study of signal transduction pathways in normal and transformed bovine T-cells
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Details
- Title
- Cloning, sequencing and expression of the bovine CD3ε and TCR-ζ chains, two invariant components of the T-cell receptor complex
- Creators
- Gerrit Hagens - Institute of Animal Pathology, University of Berne, 3012 Berne, SwitzerlandYves Galley - Institute of Animal Pathology, University of Berne, 3012 Berne, SwitzerlandIsabel Glaser - Institute of Animal Pathology, University of Berne, 3012 Berne, SwitzerlandWilliam C Davis - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA Tel. (1-509) 335-6030Cynthia L Baldwin - Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA Tel. (1-413) 545-3167Hans Clevers - Department of Immunology, Academisch Ziekenhuis Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands Tel. (31-30) 507-674Dirk A.E Dobbelaere - Institute of Animal Pathology, University of Berne, 3012 Berne, Switzerland
- Publication Details
- Gene, Vol.169(2), pp.165-171
- Academic Unit
- Veterinary Microbiology and Pathology, Department of
- Publisher
- Elsevier B.V
- Identifiers
- 99900548284701842
- Language
- English
- Resource Type
- Journal article