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From 2014.igem.org

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	This year’s project is a continuation from the previous year, using the pGAPZ(alpha) vector system we are modifying the vector to iGEM standards to allow for successful purification and secretion of recombinant DNA. Thus far, pGAPZ(alpha) A and C have been modified and we have been actively working to modify pGAPZ(alpha) B. The glyceraldehydes-3-phosphate dehydrogenase expression vector has shown high levels of protein expression in methylotrophic yeast, Pichia Pastoris. The protein is enhanced by eukaryotic posttranslational modifications that the Pichia Pastoris is capable of through methanol inducible expression. Additionally, the pGAPZ(alpha) expression system allows for controlled secretion of  a protein, making the purification of a protein relatively simple.		This year’s project is a continuation from the previous year, using the pGAPZ(alpha) vector system we are modifying the vector to iGEM standards to allow for successful purification and secretion of recombinant DNA. Thus far, pGAPZ(alpha) A and C have been modified and we have been actively working to modify pGAPZ(alpha) B. The glyceraldehydes-3-phosphate dehydrogenase expression vector has shown high levels of protein expression in methylotrophic yeast, Pichia Pastoris. The protein is enhanced by eukaryotic posttranslational modifications that the Pichia Pastoris is capable of through methanol inducible expression. Additionally, the pGAPZ(alpha) expression system allows for controlled secretion of  a protein, making the purification of a protein relatively simple.
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Revision as of 09:04, 18 August 2014

This year’s project is a continuation from the previous year, using the pGAPZ(alpha) vector system we are modifying the vector to iGEM standards to allow for successful purification and secretion of recombinant DNA. Thus far, pGAPZ(alpha) A and C have been modified and we have been actively working to modify pGAPZ(alpha) B. The glyceraldehydes-3-phosphate dehydrogenase expression vector has shown high levels of protein expression in methylotrophic yeast, Pichia Pastoris. The protein is enhanced by eukaryotic posttranslational modifications that the Pichia Pastoris is capable of through methanol inducible expression. Additionally, the pGAPZ(alpha) expression system allows for controlled secretion of a protein, making the purification of a protein relatively simple.

Using this method, the GSU iGEM team has been working to successfully insert and purify mambalgin, a protein component of the venom of Dendroaspis Polylepis, better known as Black Mamba. The mambalgin peptide is a powerful analgesic that directly blocks pain transmission in the peripheral nervous system (Diochot et al, 2012) by targeting acid sensing ion channels within nociceptors beneath the epidermis. Furthermore, recombinant purification of mambalgin could assist in developing anti-venom without the attendant risk of harvesting venom directly from snakes.