Abstract：
In this study, we present the first cloning and identification of perforin (MsPRF1) in largemouth bass (Micropterus salmoides). The full-length cDNA of MsPRF1 spans 1572 base pairs, encoding a 58.88 kDa protein consisting of 523 amino acids. Notably, the protein contains MACPF and C2 structural domains. To evaluate the expression levels of MsPRF1 in various healthy largemouth bass tissues, real-time quantitative PCR was employed, revealing the highest expression in the liver and gut. After the largemouth bass were infected by Nocardia seriolae, the mRNA levels of MsPRF1 generally increased within 48 h. Remarkably, the recombinant protein MsPRF1 exhibits inhibitory effects against both Gram-negative and Gram-positive bacteria. Additionally, the largemouth bass showed a higher survival rate in the N. seriolae challenge following the intraperitoneal injection of rMsPRF1, with observed reductions in the tissue bacterial loads. Moreover, rMsPRF1 demonstrated a significant impact on the phagocytic and bactericidal activities of largemouth bass MO/MΦ cells, concurrently upregulating the expression of pro-inflammatory factors. These results demonstrate that MsPRF1 has a potential role in the immune response of largemouth bass against N. seriolae infection.
摘要：
在这项研究中,我们介绍了大嘴鲈鱼（Micropterussalmoides）中穿孔素（MsPRF1）的首次克隆和鉴定。MsPRF1的全长cDNA跨越1572个碱基对,编码由523个氨基酸组成的58.88kDa蛋白质。值得注意的是,该蛋白质包含MACPF和C2结构域。为了评估MsPRF1在各种健康的大嘴鲈鱼组织中的表达水平，采用实时定量PCR，揭示了肝脏和肠道中最高的表达。大嘴鲈鱼被诺卡氏菌感染后，MsPRF1的mRNA水平通常在48小时内增加。值得注意的是，重组蛋白MsPRF1对革兰氏阴性和革兰氏阳性细菌均表现出抑制作用。此外,在腹膜内注射rMsPRF1后，大嘴鲈鱼在N.seriolae攻击中显示出更高的存活率，并观察到组织细菌负荷的减少。此外，rMsPRF1对大口鲈鱼MO/MΦ细胞的吞噬和杀菌活性有显著影响，同时上调促炎因子的表达。这些结果表明，MsPRF1在大口鲈鱼对抗马尾藻感染的免疫反应中具有潜在作用。
