Ruth Marian Guzman-Genuino
Institute of Biology, National Science Complex, University of the Philippines, Diliman, Quezon, 1101, Philippines
Sea anemones are inactive invertebrates that are dependent on their venom to capture and digest the prey. Moreover, this venom is also used to protect these creatures against other predators1. Their venoms are also rich in biologically active polypeptides including neurotoxins, enzymes and cytolysins2. Actinoporins are a biologically active protein which is present in the venom of sea anemones. This protein comprised of lethal pore-forming 20 kDa polypeptides.
Actinoporins have also been identified in H. crispa and are known for their hemolytic activity3. Scientists have reported cytotoxic characteristics in human tumor lines by means of p53 apoptotic pathway4.
The DNA extraction requires the use of reagent buffers with similar cell lysis properties to disrupt the cell membrane and isolate the nucleic acids of interest. However, most of these lysis buffers are responsible for causing adverse effects on human health and they are not pocket friendly.
Accordingly, new research was conducted to isolate the venom from Heteractis crispa (a sea anemone) and to analyze the presence of actinoporins and its potential to be employed as an alternate for commercial cell lysis buffers5.
For this purpose, proteome profiling of the venom extract using SDS-PAGE and silver staining was done. These steps showed the presence of proteins with molecular weights 20-215 kDa, indicating the existence of 20 kDa actinoporins among its active proteins. The venom extract was then tested for functionality as a stand-alone cell lysis reagent or as a complement to established cell lysis solutions. Heteractis crispa venom when used with sodium dodecyl sulfate as well as proteinase K, proved as effective isolation of highly pure DNA5.
Conclusively, at the end of this experiment, it was found that venom isolated from H. crispa with detergent and protease additives, is a feasible substitute for cell lysis reagent that can be efficiently employed to isolate undamaged genomic DNA with high purity.
- Sher, D., A. Knebel, T. Bsor, N. Nesher and T. Tal et al., 2005. Toxic polypeptides of the hydra-a bioinformatic approach to cnidarian allomones. Toxicon, 45: 865-879
- Linder, R. and A.W. Bernheimer, 1978. Effect on sphingomyelin-containing liposomes of phospholipase D from Corynebacterium ovis and the cytolysin from Stoichactis helianthus. Biochimica et Biophysica Acta (BBA)-Lipids Lipid Metab., 530: 236-246
- Leichenko, E.V., M.M. Monastirnaya, E.A. Zelepuga, E.S. Tkacheva and M.P. Isaeva et al., 2014. Hct-A is a New actinoporin family from the Heteractis crispa Sea anemone. Acta Nat., 6: 89-98
- Fedorov, S., S. Dyshlovoy, M. Monastyrnaya, L. Shubina and E. Leychenko et al., 2010. The anticancer effects of actinoporin RTX-A from the sea anemone Heteractis crispa (Radianthus macrodactylus). Toxicon, 55: 811-817
- Olais, J.E.N., P.Y. Luna, Z.A. Tan and R.M. Guzman-Genuino, 2015. Sebae anemone (Heteractis crispa) venom as an alternative cell lysis buffer reagent. Asian J. Cell Biol., 10: 19-24
How to cite this article:
Ruth Marian Guzman-Genuino, 2020. Heteractis crispa Venom: A Substitute for Commercial Cell Lysis Buffers. Asian Journal of Emerging Research, 2(4): 178-179.