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罗非鱼(尼罗罗非鱼)viperin表达的特征,以及通过将viperin DNA电转移到斑马鱼肌肉中来抑制细菌生长和调节免疫相关基因表达。

Characterization of tilapia (Oreochromis niloticus) viperin expression, and inhibition of bacterial growth and modulation of immune-related gene expression by electrotransfer of viperin DNA into zebrafish muscle.

作者信息

Lee Shu-Hua, Peng Kuan-Chieh, Lee Lin-Han, Pan Chieh-Yu, Hour Ai-Ling, Her Guor Mour, Hui Cho-Fat, Chen Jyh-Yih

机构信息

Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Rd., Jiaushi, Ilan 262, Taiwan.

出版信息

Vet Immunol Immunopathol. 2013 Feb 15;151(3-4):217-28. doi: 10.1016/j.vetimm.2012.11.010. Epub 2012 Nov 21.

Abstract

Viperin is an anti-viral protein, induced by viral infection. In this study, we examined whether over-expression of viperin in fish muscle could inhibit bacterial growth. We first obtained the cDNA sequence of tilapia viperin, through RT-PCR-mediated cloning and sequencing. The cDNA sequence was similar to those of several fish viperins in GenBank, and it was predicted to encode the conserved domain of radical S-adenosylmethionine superfamily proteins. Phylogenetic analysis revealed that tilapia viperin was most closely related to viperin of Sciaenops ocellatus, Coreoperca kawamebari, and C. whiteheadi. Expression of tilapia viperin was significantly up-regulated in the kidney, liver, spleen, and gills upon challenge with lipopolysaccharide (LPS) and poly(I:C) in a time- and dose-dependent manner. Injection of Vibrio vulnificus (204) and Streptococcus agalactiae (SA47) bacteria into tilapia resulted in significant induction of viperin expression in the whole body, kidney, liver, and spleen. Electrotransfer of a viperin-expressing plasmid into zebrafish muscles decreased bacterial numbers and altered expression of immune-related genes. These data indicate that such altered expression may account for the improvement in bacterial clearance following electroporation of viperin, suggesting that fish viperin has antiviral and antibacterial activities.

摘要

蝰蛇毒素蛋白是一种由病毒感染诱导产生的抗病毒蛋白。在本研究中,我们检测了鱼类肌肉中蝰蛇毒素蛋白的过表达是否能抑制细菌生长。我们首先通过逆转录聚合酶链反应(RT-PCR)介导的克隆和测序获得了罗非鱼蝰蛇毒素蛋白的cDNA序列。该cDNA序列与GenBank中几种鱼类蝰蛇毒素蛋白的序列相似,并且预测其编码自由基S-腺苷甲硫氨酸超家族蛋白的保守结构域。系统发育分析表明,罗非鱼蝰蛇毒素蛋白与眼斑拟石首鱼、河鲈和伍氏华鳊的蝰蛇毒素蛋白关系最为密切。用脂多糖(LPS)和聚肌苷酸-聚胞苷酸(poly(I:C))刺激后,罗非鱼肾脏、肝脏、脾脏和鳃中蝰蛇毒素蛋白的表达在时间和剂量依赖性上显著上调。向罗非鱼注射创伤弧菌(204)和无乳链球菌(SA47)可导致全身、肾脏、肝脏和脾脏中蝰蛇毒素蛋白表达的显著诱导。将表达蝰蛇毒素蛋白的质粒电转移到斑马鱼肌肉中可减少细菌数量并改变免疫相关基因的表达。这些数据表明,这种表达改变可能解释了蝰蛇毒素蛋白电穿孔后细菌清除率的提高,提示鱼类蝰蛇毒素蛋白具有抗病毒和抗菌活性。

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