Purification of CLoG1 from Physomitrella patens through cleaving bond of HaloⓇTag by PreScission ProteasePublic
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Physcomitrella patens (P. patens) is a eukaryotic species of moss that belongs to the family Funariaceae, from the subkingdom of bryobiotina. Since this plant performs homologous recombination efficiently, P. patens is an important resource to the genetic community. In 2018, a research article was published indicating the first discovery of CLoG1, a gene found in P. patens. The focus of our project was to determine if CLoG1 can be purified using HaloⓇ-linked resin to receive a purified product. In order to break the bond between CLoG1 and the HaloⓇ-link resin, PreScission protease (PPX) was needed for cleavage. His-PPX and GST-Snc2 were both successfully purified via their respective tags, and gel electrophoresis confirmed these results. Once both were successfully purified, PPX effectively cleaved the bond between GST-Snc2 proving PPX works. After analyzing with a gel and immunoblot, the purification of CLoG1 was inconclusive, but research is still ongoing into the purification of GFP through HaloⓇ-Link resin.
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