Jellyfish bioactive compounds: Methods for wet-lab work
2016
Frazão B. | Antunes A. | CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental
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. © 2016 by the authors; licensee MDPI, Basel, Switzerland.
显示更多 [+] 显示较少 [-]We are thankful to the Academic Editor and the two anonymous reviewers for their valuable comments and suggestions. B.F. was funded by a PhD grant (SFRH/BD/48184/2008) by Fundação para a Ciência e a Tecnologia (FCT). A.A. was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT and European Regional Development Fund (ERDF) in the framework of the programme PT2020 and the FCT project PTDC/AAG-GLO/6887/2014.
显示更多 [+] 显示较少 [-]