Acta Scientiarum Naturalium Universitatis Sunyatseni ›› 2020, Vol. 59 ›› Issue (5): 144-155.doi: 10.13471/j.cnki.acta.snus.2020.01.09.2020E002

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Cloning and expression analysis of Nile tilapia TNF-R1 and TNF-R2 gene

YE HangyuGE YanyanWU JinyingZHAN Xuliang   

  1. School of Life Sciences,Sun Yat-sen University/Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals/Guangdong Provincial Engineering Technology Research Center for Healthy Breeding of Important Economic FishGuangzhou 510275China
  • Received:2020-01-09 Online:2020-09-25 Published:2020-09-26

Abstract: To reveal the role of TNF-α receptor genes TNF-R1 and TNF-R2 in the Nile tilapis(Oreochromis niloticusimmune processeswe cloned the full-length cDNA sequences of TNF-R1 and TNFR2 from Nile tilapia by RACE. Sequence analysis showed that the full length of TNF-R1 cDNA was 2 275 bpcontaining an open reading frame of 1 335 bpand encoding a protein with 444 amino acidsthe full length of TNF-R2 cDNA was 1 719 bpcontaining an open reading frame of 1 476 bpand encoding a protein with 491 amino acids. Protein structure prediction showed that TNF-R1 have three cysteinerich domains and an intracellular death domainwhile TNF-R2 have four cysteine-rich domains and without death domain. Systematic evolutionary analysis showed that the tilapia TNF-R1 and TNF-R2 were highly homologous with those of other vertebrates. Phylogenetic tree showed that TNF-R1and TNF-R2 were clustered separately. By qRT-PCRthe highest expression levels of TNF-R1 and TNF-R2 mRNA were found in the gillhead kidney and spleenand the lowest levels were in pituitary and muscle. The expression of TNF-α,TNF-R1 and TNF-R2 genes can be increased to varying degrees by stimulating the tilapia kidneys and spleen white cells incubated with LPS and Poly ⅠCrespectively. The expressions of three genes in the headkidney and spleen can be increased after intra-peritoneal injection of Streptococcus agalactiae. This study has some reference value to show the immune regulation mechanisms of TNF-R1and TNF-R2 to improve the level of healthy breeding of tilapia.

Key words: TNF-α, TNF-R1, TNF-R2, Oreochromis niloticus, immunity, Streptococcus agalactiae

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