Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5905412 | Gene | 2015 | 8 Pages |
Abstract
CD3 is one of the most important cell surface markers of T lymphocytes which play an important role in signal transmission of antigen recognition. In this study, goose CD3ε gene was cloned by touchdown PCR with the template of goose thymus cDNA. The complete open reading frame of goose CD3ε encoded 178 amino acid residues with a 21 signal peptide. Sequence alignments showed that goose CD3ε had an amino acid sequence similarity to duck (80.3%) and chicken (66.3%). The extracellular domain of goose CD3ε was efficiently expressed as fusion protein in Escherichia coli, purified by a Ni-NTA agarose column, and the purified recombinant protein was used to produce anti-GoCD3εex polyclonal antibodies. The characteristics of PAb were identified by Western blot, cellular ELISA, IFA, FCM, and LSCM analysis. These results may be useful for a better understanding of goose CD3ε and have a foundation for the study of T cell mediated immune mechanism in waterfowl.
Keywords
ITAMsIFABRSHRPTCrPBLsDAPIIPTGLSCMFITCPBMCmAbCD3εFCMPAB4′,6-diamidino-2-phenylindolecDNAComplementary DNAindirect immunofluorescence assayMonoclonal antibodyPolyclonal antibodyenzyme linked immunosorbent assayExpressionisopropyl β-D-1-thiogalactopyranosideELISACharacterizationFlow cytometricimmunoreceptor tyrosine-based activation motifsGoosefluorescein isothiocyanateperipheral blood lymphocytesmajor histocompatibility complexMHCMolecular cloningHorseradish peroxidasePropidium iodideT-cell receptor
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Genetics
Authors
Xuelian Zhang, Shuangshi Wei, Jianwei Shao, Shudong Zhang, Mingchun Gao, Wenlong Zhang, Bo Ma, Junwei Wang,