Department of Biochemistry & Molecular Biology
نویسنده
چکیده
Activation-Induced Deaminase (AID) protein functions in class switch recombination and somatic hypermutation in mammalian cells. The AID protein deaminates cytidine sites in single stranded DNA. We designed and constructed an expression plasmid that encodes AID as a recombinant fusion protein. Our construct includes an N-terminal maltose binding protein as a solubility and expression tag followed by a hexa-histidine tag to allow affinity purification linked via a TEV cleavage site to AID. This construct will allow us to express the fusion protein at adequate levels, to purify the fusion protein using NiNTA affinity chromatography, to cleave the fusion protein with TEV protease releasing native AID, and to purify the native AID away from both the his-tagged TEV protease as well as the cleaved N-terminal tag using a second NiNTA affinity chromatography step. In previous work with recombinant AID protein, it was found that amino acids remaining after tag cleavage altered AID function. As our new construct will allow us to produce AID protein with no non-native amino acids we expect that it will have full wild type function. “THE ROLE OF PHOTOPERIOD IN COLD ACCLIMATION” Cynthia Collings, under the direction of Dr. Michael F. Thomashow, Plant Research Laboratory
منابع مشابه
Genetic diversity analysis of aquaculture strains of Acipenser stellatus (Pallas, 1771) using DNA markers
Acipenser stellatus (stellate sturgeon) represents a sturgeon species of great importance for the scientific community and fishermen that has been intensively captured for decades. Besides of overfishing and poaching, their reproduction sites have been destroyed and their natural habitats were affected by pollution. In consequence, wild populations of stellate sturgeon have decreased dramatical...
متن کاملGenetic diversity analysis of aquaculture strains of Acipenser stellatus (Pallas, 1771) using DNA markers
Acipenser stellatus (stellate sturgeon) that has been intensively captured for decades represents a sturgeon species of great importance for the scientific community and fishermen. Besides overfishing and poaching, their reproduction sites have been destroyed and their natural habitats were affected by pollution. In consequence, wild populations of stellate sturgeon have decreased dramatically ...
متن کاملThe 15th Iranian National Congress of Biochemistry & 6th International Congress of Biochemistry and Molecular Biology
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The downregulation of ATG4B mediated by microRNA-34a/34c-5p suppresses rapamycin-induced autophagy
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