Functional Investigations of Proteins and Enzymatic Toxins from Full-Length-Enriched cDNA of Timber Rattlesnake (Crotalus Horridus)
Snake Venom is a highly modified form of saliva, contains hundreds of zootoxins, necrotoxins, cytotoxins, neurotoxins, and mytotoxins, all of which are primarily made up of mostly proteins, polypeptides, and other components such as enzymes, lipids, and carbohydrates. Although snake venom is known f...
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JSU Digital Commons
2020
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| Online Access: | https://digitalcommons.jsu.edu/etds_theses/2 https://digitalcommons.jsu.edu/cgi/viewcontent.cgi?article=1001&context=etds_theses |
| Summary: | Snake Venom is a highly modified form of saliva, contains hundreds of zootoxins, necrotoxins, cytotoxins, neurotoxins, and mytotoxins, all of which are primarily made up of mostly proteins, polypeptides, and other components such as enzymes, lipids, and carbohydrates. Although snake venom is known for its harmful effects, it has also had a positive impact in scientific discovery and medicine. Some toxins from snakes function to inhibit pain, and this means they can be very effective analgesics. For instance, crotalphine is considered a 14-amino-acid-peptide that has a disulfide bond and shows analgesic properties through TRPA1 desensitization. This is the reason why snake venom peptides are at our center of interest; there are a plethora of new compounds to discover. Venomous snakes represent a large pool of potentially bioactive proteins, and despite recent computational studies and discovery efforts of over 2200 sequences, most of the snake venom toxins’ functions are still uncharacterized. Using RNA pooled from the venom gland tissues of our sample, we constructed a cDNA library. In order to generate full-length transcripts, we used the SMARTTM technique for first-strand cDNA synthesis combined with duplex-specific nuclease enzyme from Kamchatka crab for cDNA normalization. Five hundred plaques were successfully screened, and 50 of them were sequenced. The plaques that we screened, amplified, and analyzed using agarose gel electrophoresis and then sequenced mostly coded for ribosomal proteins. Crotamine, C-type lectin (CLEC), phospholipase (PLA2), and serine protease were the components we were able to identify when we proceeded to annotate our 4 selected sequences with the venom-associated genes. Using predictive modeling, we determined specific locations, coding regions, variation information, exons, introns, and functions. Crotamine functions to inhibit pain, and this means it can be a very effective analgesic; it is a cell-penetrating peptide and a suitable means for transporting macromolecules through ... |
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