Carriero Mateus Maldonado, Henrique-Silva Flavio, Meira Caroline Munhoz, Gato Igor Mateus Queiroz, Caetano Alexandre Rodrigues, Lobo Francisco Pereira, Alves Anderson Luis, Varela Eduardo Sousa, Maia Antonio Augusto Mendes
Universidade de São Paulo, Faculdade de Zootecnia e Engenharia de Alimentos, Departamento de Medicina Veterinária, Pirassununga, SP, Brazil.
Universidade Federal de São Carlos, Departamento de Genética e Evolução, São Carlos, SP, Brazil.
Genet Mol Biol. 2020 Nov 6;43(4):e20200006. doi: 10.1590/1678-4685-GMB-2020-0006. eCollection 2020.
In the present study, the complete characterization of cDNA and genomic sequences of IL-1β and IL-8, as well as the expression profile of these genes in the South American fish pacu (Piaractus mesopotamicus) is provided. The full-length pmIL-1β cDNA was composed of 1208 nucleotides that would produce a precursor peptide with 273 amino acid residues. A putative caspase-1 cleavage site, similar to what is found in mammalian IL-1β, was identified producing a mature peptide with a theoretical molecular weight of 17.21 kDa. The pmIL-8 cDNA sequence consisted of 1019 nucleotides which encoded a 95-amino acid protein with a theoretical molecular weight of 10.43 kDa that showed all typical CXC chemokine features, including a 20-residue signal peptide and four conserved cysteine residues. Constitutive mRNA expression was detected for both genes in the liver, head kidney, gill, intestine, skin and spleen. After a bacterial challenge, up-regulation was detected for both pmIL-1β and pmIL-8 in the spleen and head kidney at 12 h post-infection. At 24 h post-infection there was a decrease in the expression of both genes, with pmIL-8 showing a significant down-regulation in the liver and head kidney when compared to the control groups.
在本研究中,提供了南美脂鲤(Piaractus mesopotamicus)中白细胞介素-1β(IL-1β)和白细胞介素-8(IL-8)的cDNA和基因组序列的完整特征,以及这些基因的表达谱。全长pmIL-1β cDNA由1208个核苷酸组成,可产生具有273个氨基酸残基的前体肽。鉴定出一个类似于哺乳动物IL-1β中的半胱天冬酶-1切割位点,产生理论分子量为17.21 kDa的成熟肽。pmIL-8 cDNA序列由1019个核苷酸组成,编码一种95个氨基酸的蛋白质,理论分子量为10.43 kDa,具有所有典型的CXC趋化因子特征,包括一个20个残基的信号肽和四个保守的半胱氨酸残基。在肝脏、头肾、鳃、肠道、皮肤和脾脏中均检测到这两个基因的组成型mRNA表达。细菌攻击后,感染后12小时在脾脏和头肾中检测到pmIL-1β和pmIL-8均上调。感染后24小时,这两个基因的表达均下降,与对照组相比,pmIL-8在肝脏和头肾中表现出显著下调。