Modification of Aminoacyl tRNA Synthetase in Order to Incorporate an Unnatural Amino Acid

Proteins allow daily processes in the cell to occur. A protein consists of amino acids. There are twenty natural amino acids coded for in the DNA of organisms. The natural amino acids can be modified to form unnatural amino acids (UAAs). UAAs have useful characteristics when inserted into a protein of a cell, like the ability of fluoresce, which makes their incorporation important in research. For an UAA to be incorporated into a protein, it must be bound to a transport RNA molecule by an enzyme called aminoacyl tRNA synthetase (aaRS). An existing aaRS was modified in E. Coli bacterial cells to incorporate 3-(2-pyridyl)-L-Alanine since it has metal-binding capabilities. Once incorporated, the UAA acts as a sensor for a metal, making it useful to environmental fields. The aaRS was mutated using saturation mutagenesis at sites L32, V65, W108, G158, A159. The cells were run through a positive screen to determine if the mutated aaRS incorporated the UAA into a green fluorescent protein, which glowed if the UAA was inserted. The results of the positive screen showed mutated aaRSs 2, 4, 7, and 8 incorporated 3-(2-pyridyl)-L-Alanine, while mutated aaRSs 2, 5, 6, 7, 8, and 9 incorporated p-cyanophenylalanine. A negative screen to test if the mutated aaRS only incorporate an UAA, not natural amino acids still present in the cell, will be run on the mutated aaRSs passing the positive screen

Modification of Aminoacyl tRNA Synthetase in Order to Incorporate an Unnatural Amino Acid

Proteins allow daily processes in the cell to occur. A protein consists of amino acids. There are twenty natural amino acids coded for in the DNA of organisms. The natural amino acids can be modified to form unnatural amino acids (UAAs). UAAs have useful characteristics when inserted into a protein of a cell, like the ability of fluoresce, which makes their incorporation important in research. For an UAA to be incorporated into a protein, it must be bound to a transport RNA molecule by an enzyme called aminoacyl tRNA synthetase (aaRS). An existing aaRS was modified in E. Coli bacterial cells to incorporate 3-(2-pyridyl)-L-Alanine since it has metal-binding capabilities. Once incorporated, the UAA acts as a sensor for a metal, making it useful to environmental fields. The aaRS was mutated using saturation mutagenesis at sites L32, V65, W108, G158, A159. The cells were run through a positive screen to determine if the mutated aaRS incorporated the UAA into a green fluorescent protein, which glowed if the UAA was inserted. The results of the positive screen showed mutated aaRSs 2, 4, 7, and 8 incorporated 3-(2-pyridyl)-L-Alanine, while mutated aaRSs 2, 5, 6, 7, 8, and 9 incorporated p-cyanophenylalanine. A negative screen to test if the mutated aaRS only incorporate an UAA, not natural amino acids still present in the cell, will be run on the mutated aaRSs passing the positive screen

Synthetic Routes to a Library of Novel Methionine Synthase Inhibitors

Fungal infections are of continuous concern, especially with regard to immunocompromised patients. In an effort to develop new potential anti-fungal agents, we have begun synthesizing a library of potential inhibitors of the fungal Methionine Synthase (MetSyn) enzyme. Key differences between the B12-independant fungal MetSyn enzyme and the B12-dependant mammalian form can allow for an antifungal drug to be developed to exclusively bind the fungal enzyme and inhibit fungal growth while leaving the host (patient) unaffected. We are currently exploring the synthesis of various pterin and deazaguanine-based molecules as these mimic folate, an essential substrate for MetSyn function. We have begun testing these new molecules for activity in a fungal growth assay, as well as a fluorescent assay for monitoring MetSyn activity

Synthetic Routes to a Library of Novel Methionine Synthase Inhibitors

Fungal infections are of continuous concern, especially with regard to immunocompromised patients. In an effort to develop new potential anti-fungal agents, we have begun synthesizing a library of potential inhibitors of the fungal Methionine Synthase (MetSyn) enzyme. Key differences between the B12-independant fungal MetSyn enzyme and the B12-dependant mammalian form can allow for an antifungal drug to be developed to exclusively bind the fungal enzyme and inhibit fungal growth while leaving the host (patient) unaffected. We are currently exploring the synthesis of various pterin and deazaguanine-based molecules as these mimic folate, an essential substrate for MetSyn function. We have begun testing these new molecules for activity in a fungal growth assay, as well as a fluorescent assay for monitoring MetSyn activity

Investigation of the Parasympathetic Effects of Lavender Essential Oil in Humans

The purpose of this study will be to investigate the claim that administration of lavender (Lavandula angustifolia) essential oil (topically, orally, and/or respiratorily) produces a relaxative effect in human subjects. This investigation will theoretically be conducted in two stages. Stage one will focus primarily on determining the presence of therapeutic effects and the relative effectiveness of lavender in several application modalities. Stage two will proceed based on findings from stage one. If significant parasympathetic effects are observed in relation to one or more of the lavender oil modalities described above, a more focused investigation will be conducted in stage two to ascertain the specific active chemical component(s) in the oil that stimulate(s) the therapeutic effect

Pacific Portraits: The People Behind the Scenes at Pacific University (Volume Three)

There\u27s a saying that if you build a fire on a hill, all sorts of interesting people will climb that hill to meet you, to hear your story or to tell one, or both. In these pages, the students brought the fire to a variety of locations on and off campus to meet interesting characters, staff some of us see on a daily basis but hardly know a thing about. We invite you to peer into these fires, once again, and gaze at the vivid lives of members of our Pacific community to hear a good story, and perhaps, inspire your own.https://commons.pacificu.edu/beetree/1008/thumbnail.jp

Modification of Aminoacyl tRNA Synthetase in Order to Incorporate An Unnatural Amino Acid

Proteins allow daily processes in the cell to occur. A protein consists of amino acids. There are twenty natural amino acids coded for in the DNA of organisms. The natural amino acids can be modified to form unnatural amino acids (UAAs). UAAs have useful characteristics when inserted into a protein of a cell, like the ability of fluorescence and the ability to undergo unique reactions. For an UAA to be incorporated into a protein, it must be bound to a transport RNA molecule by an enzyme called aminoacyl tRNA synthetase (aaRS). An existing aaRS was modified in E. Coli bacterial cells to incorporate 3-(2-pyridyl)-L-Alanine since it has metal-binding capabilities. Once incorporated, the UAA could act as a sensor for a metal, making it useful to environmental fields. The aaRS was randomly mutated using saturation mutagenesis at sites L32, V65, W108, G158, A159. The cells were run through a positive screen to determine if the mutated aaRS incorporated the UAA into a green fluorescent protein, which glowed if the UAA was inserted. The results of the positive screen showed several mutated aaRSs (2,4,7,8) incorporated 3-(2-pyridyl)-L-Alanine, while other mutants (2,5,6,7,8,9) also/instead incorporated the p-cyanophenylalanine amino acid that the original aaRS was designed to select

Modification of Aminoacyl tRNA Synthetase in Order to Incorporate An Unnatural Amino Acid

Proteins allow daily processes in the cell to occur. A protein consists of amino acids. There are twenty natural amino acids coded for in the DNA of organisms. The natural amino acids can be modified to form unnatural amino acids (UAAs). UAAs have useful characteristics when inserted into a protein of a cell, like the ability of fluorescence and the ability to undergo unique reactions. For an UAA to be incorporated into a protein, it must be bound to a transport RNA molecule by an enzyme called aminoacyl tRNA synthetase (aaRS). An existing aaRS was modified in E. Coli bacterial cells to incorporate 3-(2-pyridyl)-L-Alanine since it has metal-binding capabilities. Once incorporated, the UAA could act as a sensor for a metal, making it useful to environmental fields. The aaRS was randomly mutated using saturation mutagenesis at sites L32, V65, W108, G158, A159. The cells were run through a positive screen to determine if the mutated aaRS incorporated the UAA into a green fluorescent protein, which glowed if the UAA was inserted. The results of the positive screen showed several mutated aaRSs (2,4,7,8) incorporated 3-(2-pyridyl)-L-Alanine, while other mutants (2,5,6,7,8,9) also/instead incorporated the p-cyanophenylalanine amino acid that the original aaRS was designed to select

The Philomathean

Monthly newspaper from Chappell Hill, Texas that includes local news and commentary related to the Chappell Hill Female College

The Philomathean

Monthly newspaper from Chappell Hill, Texas that includes local news and commentary related to the Chappell Hill Female College