Sebae Anemone (Heteractis crispa) Venom as an Alternative Cell Lysis Buffer Reagent

摘要

Sea anemone venoms are rich reservoirs of biologically active proteins which include actinoporins, a large family of lethal pore-forming 20 kDa polypeptides. In this study, venom isolated from leathery sebae anemone Heteractis crispa, was examined forpresence of actinoporins and its potential to be used as a substitute for commercial cell lysis buffers. Proteome profiling of the venom extract using SDS-PAGE and silver staining revealed proteins with molecular weights 20-215 kDa, indicating the presence of 20 kDa actinoporins among its active proteins. The venom extract was 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 sulfateand proteinase K, demonstrated efficient isolation of highly pure DNA. The isolated nucleic acid samples were run in 1% (w/v) agarose gel and confirmed the absence of nuclease activity throughout all treatments. Taken together, venom isolated from H. crispa with detergent and protease additives, was found to be a viable alternative cell lysis reagent used in isolating undamaged genomic DNA with high purity.