Identification and functional analysis of the perforin-1 like gene in disease resistance in half smooth tongue sole (Cynoglossus semilaevis)

文献类型： 外文期刊

第一作者： Fu, Xiaoqin

作者： Fu, Xiaoqin;Zhao, Fazhen;Chen, Songlin;Fu, Xiaoqin;Chen, Yadong;Wang, Lei;Zhou, Qian;Li, Ming;Song, Yu;Li, Yangzhen;Zhao, Fazhen;Chen, Songlin;Fu, Xiaoqin;Chen, Yadong;Wang, Lei;Zhou, Qian;Li, Ming;Song, Yu;Li, Yangzhen;Zhao, Fazhen;Chen, Songlin;Fu, Xiaoqin;Chen, Yadong;Wang, Lei;Zhou, Qian;Li, Ming;Song, Yu;Li, Yangzhen;Zhao, Fazhen;Chen, Songlin

作者机构：

关键词： Cynoglossus semilaevis; Vibrio harveyi; GWAS; rCsPRF1l; Immune defense

期刊名称：DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY （ 影响因子：3.636； 五年影响因子：3.654 ）

ISSN： 0145-305X

年卷期： 2021 年 122 卷

页码：

收录情况： SCI

摘要： The pore-forming protein perforin is one of the effectors of cell-mediated killing via the granule exocytosis pathway. In this study, a genome-wide association study was conducted in Vibrio harveyi disease-resistant and disease-susceptible families of half smooth tongue sole (Cynoglossus semilaevis) to determine the genes accounting for host resistance, and a perforin homologue was identified, designated perforin-1 like (CsPRF1l). The full-length cDNA of CsPRF1l is 1835 bp, and encodes 514 amino acids. The CsPRF1l gene consists of 10 exons and 9 introns, spanning approximately 7 kb. The amino acid sequence of CsPRF1l shows 60.35, 54.03, 41.92, and 34.17% identities to Morone saxatilis PRF1l, Oryzias melastigma PRF1l, Danio rerio PRF1.5 and Homo sapiens PRF, respectively. Sequence analysis revealed the presence of membrane attack complex/perforin (MACPF) and C2 domains in CsPRF1l. Quantitative real-time PCR showed that CsPRF1l presented a higher intestinal expression level in disease-resistant families than in susceptible families. Tissue expression pattern analysis showed that CsPRF1l is present in most of the tested tissues and highly expressed in the intestine, brain, stomach and gills. After challenge with V. harveyi, CsPRF1l mRNA was markedly upregulated in the liver, spleen, kidney, intestine, gills and skin. In addition, the recombinant CsPRF1l protein exhibited obvious antimicrobial activity against V. harveyi in vitro and in a zebrafish model. Collectively, these data indicate that CsPRF1l modulates host immune defense against V. harveyi invasion and provide clues about the efficacy of rCsPRF1l in fish that will give rise to useful therapeutic applications for V. harveyi infection in C. semilaevis.

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