Frazão Bárbara, Antunes Agostinho
CIIMAR/CIMAR, Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas 177, 4050-123 Porto, Portugal.
Department of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal.
Mar Drugs. 2016 Apr 12;14(4):75. doi: 10.3390/md14040075.
The study of bioactive compounds from marine animals has provided, over time, an endless source of interesting molecules. Jellyfish are commonly targets of study due to their toxic proteins. However, there is a gap in reviewing successful wet-lab methods employed in these animals, which compromises the fast progress in the detection of related biomolecules. Here, we provide a compilation of the most effective wet-lab methodologies for jellyfish venom extraction prior to proteomic analysis-separation, identification and toxicity assays. This includes SDS-PAGE, 2DE, gel chromatography, HPLC, DEAE, LC-MS, MALDI, Western blot, hemolytic assay, antimicrobial assay and protease activity assay. For a more comprehensive approach, jellyfish toxicity studies should further consider transcriptome sequencing. We reviewed such methodologies and other genomic techniques used prior to the deep sequencing of transcripts, including RNA extraction, construction of cDNA libraries and RACE. Overall, we provide an overview of the most promising methods and their successful implementation for optimizing time and effort when studying jellyfish.
随着时间的推移，对海洋动物生物活性化合物的研究提供了源源不断的有趣分子来源。水母因其有毒蛋白质而成为常见的研究对象。然而，在综述这些动物所采用的成功的湿实验室方法方面存在差距，这不利于相关生物分子检测的快速进展。在此，我们汇编了蛋白质组学分析前用于水母毒液提取的最有效的湿实验室方法——分离、鉴定和毒性测定。这包括SDS-PAGE、双向电泳、凝胶色谱、高效液相色谱、二乙氨基乙基纤维素、液相色谱-质谱联用、基质辅助激光解吸电离、蛋白质印迹、溶血试验、抗菌试验和蛋白酶活性试验。为了采用更全面的方法，水母毒性研究应进一步考虑转录组测序。我们综述了在转录本深度测序之前使用的此类方法和其他基因组技术，包括RNA提取、cDNA文库构建和cDNA末端快速扩增。总体而言，我们概述了最有前景的方法及其在研究水母时优化时间和精力的成功应用。
