Molecular cloning, characterization and expression analysis of A cDNA encoding T cell receptor beta gene in nile tilapia (Oreochromis niloticus, Linn.) | |
| Author | Nudtha Nithikulworawong |
| Call Number | AIT Diss. no.AQ-12-01 |
| Subject(s) | Nile tilapia Molecular cloning T-cells--Receptors |
| Note | A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Technical Science |
| Publisher | Asian Institute of Technology |
| Series Statement | Dissertation ; no. AQ-12-01 |
| Abstract | T cell receptors (TCRs) are fundamental mediators of acquire immune responses as TRC αβ on T cells recognize foreign structures (peptides derived from processed antigens) bound to the major histocompatibility (HMC) on antigen presenting cells (APC). Hence, the complete of TCR β cDNA sequences from Nile tilapia (Oreochromis niloticus), which is an important commercial species for aquaculture in Thailand, was cloned by 5' rapid Amplification Complementary DNA Ends (RACE) technique in this study. The full-length cDNA sequence of TCR β chain of Nile tilapia composed of 1,263-bp. This complete sequence contained 35-bp of 5' untranslated regions (UTR), 283-bp of 3' UTR and a 942-bp open reading frame (ORF) encoding a 314-amino-acid protein. A multiple alignment of Nile tilapia's TCR β chain which other known TCR β sequences showed that conserved four cysteine residues involved in the disulfide bonds formation and some conserved amino acids supposed to be important for gathering and signaling of the TCR αβ/CD3 complex, both of which are normally found in the TCR β chain of other vertebrates. Amino acid and nucleotide sequence alignment indicated that Nile tilapia TCR β was closely similar to gilthead seabream (Sparus aurata) and bicolor damselfish (Stegastes partitus). Phylogenetic analysis strongly supported that Nile tilapia TCR β was clustered in the group of teleost TCR β chain. Using semi-quantitative and quantitative RT-PCP, the highest expression level of TCR β was detected in the thymus. Interestingly, Streptococcus agalactiae significantly induced up-regulation of the TCR β chain, and the strongest up-regulation was detected in the brain and peripheral blood leukocytes (PBLs). In in vitro experiments, concanavalin A and Aeromonas hydrophila were found to be significantly increased the expression of the TCR β chain in PBLs after 48 h (p<0.01) and 72 h (p<0.05), respectively. Furthermore, quantitative real-time PCR analysis showed that intraperitoneal injection (IP) of 10 cfu/mL of S. agalactiae could induce TCR β expression greater than the administration of 10 cfu/mL. The presence of the TCR β chain observed in fish in this study suggests the presence of T cell populations that have been found in higher vertebrates may play a crucial functional role in fish to response to pathogens |
| Year | 2012 |
| Corresponding Series Added Entry | Asian Institute of Technology. Dissertation ; no. AQ-12-01 |
| Type | Dissertation |
| School | School of Environment, Resources, and Development (SERD) |
| Department | Department of Food, Agriculture and Natural Resources (Former title: Department of Food Agriculture, and BioResources (DFAB)) |
| Academic Program/FoS | Aquaculture and Aquatic Resources Management (AQ) |
| Chairperson(s) | Yakupitiyage, Amararatne |
| Examination Committee(s) | Gallardo, Wenresti G.;Anal, Anil Kumar;Prapansak Srisapoome;Singh, I. S. Bright |
| Scholarship Donor(s) | Commission on Higher Education;Royal Thai Government |
| Degree | Thesis (Ph.D.) - Asian Institute of Technology, 2012 |