Molecular cloning and expression of feline CD3epsilon
| Autor: | Y, Nishimura, T, Miyazawa, Y, Ikeda, Y, Izumiya, K, Nakamura, J S, Cai, E, Sato, M, Kohmoto, T, Mikami |
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| Rok vydání: | 1998 |
| Předmět: |
Base Sequence
CD3 Complex Sequence Homology Amino Acid Reverse Transcriptase Polymerase Chain Reaction Blotting Western Molecular Sequence Data Sequence Analysis DNA Transfection Molecular Weight Open Reading Frames Gene Expression Regulation Cats Animals Humans Amino Acid Sequence RNA Messenger Cloning Molecular Fluorescent Antibody Technique Indirect Baculoviridae Sequence Alignment DNA Primers |
| Zdroj: | Veterinary immunology and immunopathology. 65(1) |
| ISSN: | 0165-2427 |
| Popis: | The cDNA of feline CD3epsilon, one of the T-cell receptor components, was cloned from a feline T-lymphoblastoid cell line (MYA-1 cells) and peripheral blood mononuclear cells and thymocytes of cats by polymerase chain reaction. Sequencing analysis revealed that the open reading frame of feline CD3epsilon consists of 606 base pairs encoding a predicted molecular mass of 25 kDa transmembrane protein which lacks N-glycosylation site. Comparison of the predicted amino acid sequence of feline CD3epsilon with those of other mammalians' homologues revealed that a relatively low homology was present in the extracellular domain. However, the cytoplasmic domain contained several characteristic motifs highly conserved across the species. These motifs were known to be important for signal transduction upon T-cell activation or endoplasmic reticulum retention. In addition, the feline CD3epsilon protein was expressed in an insect cell line (Sf9) by a baculovirus expression system. The expression was confirmed by indirect immunofluorescence assay and immunoblotting analysis using an anti-human CD3epsilon polyclonal antibody. These results will provide additional information for understanding the feline immune system. |
| Databáze: | OpenAIRE |
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