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Functional characterization of NK-lysin in golden pompano Trachinotus ovatus (Linnaeus 1758)

文献类型： 外文期刊

作者： Liu, Bo 1 ; Liu, Guang-Dong 1 ; Guo, Hua-Yang 1 ; Zhu, Ke-Cheng 1 ; Guo, Liang 1 ; Liu, Bao-Suo 1 ; Zhang, Nan 1 ; Jiang, 1 ;

作者机构： 1.Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Key Lab South China Sea Fishery Resources Exploit, Minist Agr & Rural Affairs, Guangzhou 510300, Guangdong, Peoples R China

2.Guangdong Prov Engineer Technol Res Ctr Marine Bi, Guangzhou, Guangdong, Peoples R China

3.Guangdong Prov Key Lab Fishery Ecol & Environm, Guangzhou, Guangdong, Peoples R China

关键词： Antibacterial activity; Antimicrobial peptide; Trachinotus ovatus; NK-Lysin; Photobacterium damselae

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： 2020 年 107 卷

页码：

收录情况： SCI

摘要： NK-lysin is an important part of the innate immune defence system and plays an important role in resisting the invasion of pathogenic microorganisms. In this study, NK-lysin from golden pompano (Trachinotus ovatus) was characterized and its expression in response to Photobacterium damselae was investigated. The full-length NKlysin cDNA was 731 bp, which comprised a 5'-UTR of 63 bp, an ORF of 444 bp, and a 3'-UTR of 224 bp, and encoded 147 amino acids; NK-lysin consisted of a conserved saposin B domain and six conserved cysteines that formed three pairs of disulfide bonds. The genomic organization of NK-lysin was also determined and the gene consisted of four introns and five exons. The predicted promoter region of ToNK-lysin contained several putative transcription factor binding sites. Quantitative real-time (qRT-PCR) analysis indicated that ToNK-lysin was ubiquitously expressed in all examined tissues; the highest mRNA levels were observed in the skin, kidney and intestine, while the lowest expression level was detected in the stomach. After P. damselae stimulation, the expression level of NK-lysin mRNA was significantly upregulated in various tissues of golden pompano. In addition, SDS-PAGE showed that the molecular mass of recombinant NK-lysin expressed in pGEX-6P-1 was approximately 37 kDa. The purified recombinant protein showed antibacterial activity against gram-positive and gram-negative bacteria. The results indicate that golden pompano NK-lysin has potential antimicrobial roles in fish innate immunity.

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