{"id":{"repo_id":"unr","oai_identifier":"oai:scholarwolf.unr.edu:11714/641"},"canonical_url":"https://search.dev.ndltd.org/etd/unr/oai:scholarwolf.unr.edu:11714/641","repository":{"repo_id":"unr","name":"University of Nevada - Reno","base_url":"https://scholarwolf.unr.edu/server/oai/request"},"display":{"title":"Resin Detoxification in Dendroctonus ponderosae: Functional Characterizations of Cytochrome P450 Proteins","abstract":"A growing epidemic of pine tree deforestation in North America can largely be attributed to a particular pest, the Mountain Pine Beetle (MPB, Dendroctonus ponderosae). Coupling pheromone-coordinated mass attacks with an ability to resist the trees' natural defenses allows the beetles to overwhelm their targets and burrow beneath the tree bark to reproduce. MPB use cytochrome P450 enzymes to both detoxify toxic monoterpene compounds present in tree resin as well as synthesize the pheromones used during tree invasion, suggesting an evolutionary link between the P450 proteins involved in the two processes. This study attempts to contribute to the understanding of this evolutionary link by examining CYP6DH3, a P450 closely related to a known MPB resin detoxifier CYP6DH2. To do so, microsomal CYP6DH3 was expressed in Sf9 insect cells before being separated and incubated with various monoterpene substrates. GC screening revealed that no products could consistently be formed, which suggested an issue with the assay protocol. Various adjustments to the procedure have allowed the results of a previous study (Song et al., 2013) to be replicated, though further testing must be done to confirm whether these reaction conditions apply to CYP6DH3.","abstract_html":"A growing epidemic of pine tree deforestation in North America can largely be attributed to a particular pest, the Mountain Pine Beetle (MPB, Dendroctonus ponderosae). Coupling pheromone-coordinated mass attacks with an ability to resist the trees' natural defenses allows the beetles to overwhelm their targets and burrow beneath the tree bark to reproduce. MPB use cytochrome P450 enzymes to both detoxify toxic monoterpene compounds present in tree resin as well as synthesize the pheromones used during tree invasion, suggesting an evolutionary link between the P450 proteins involved in the two processes. This study attempts to contribute to the understanding of this evolutionary link by examining CYP6DH3, a P450 closely related to a known MPB resin detoxifier CYP6DH2. To do so, microsomal CYP6DH3 was expressed in Sf9 insect cells before being separated and incubated with various monoterpene substrates. GC screening revealed that no products could consistently be formed, which suggested an issue with the assay protocol. Various adjustments to the procedure have allowed the results of a previous study (Song et al., 2013) to be replicated, though further testing must be done to confirm whether these reaction conditions apply to CYP6DH3.","abstract_has_math":false,"creators":["Bernardo, Joseph"],"institution":"University of Nevada, Reno","degree_name":"Biochem and Molecular Biology","degree_level":"Honors Thesis","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Tittiger, Claus R."],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015","date_published":"2015","updated_at":"2026-07-27T21:46:08Z","subjects":[],"languages":["en_US","English"],"rights":["In Copyright(All Rights Reserved)"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/11714/641","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Tittiger, Claus R."]},{"key":"dc:creator","label":"Author","values":["Bernardo, Joseph"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2017-01-24T23:09:18Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2017-01-24T23:09:18Z"]},{"key":"dc:date.issued","label":"Date","values":["2015"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Honors Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Biochem and Molecular Biology"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Nevada, Reno"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["English"]},{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]},{"key":"dc:rights","label":"Dc Rights","values":["In Copyright(All Rights Reserved)"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/11714/641"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The University of Nevada, Reno Libraries will promptly respond to removal requests related to content that violates intellectual property laws, data protections, or has been uploaded without creator consent. Takedown notices should be directed to our ScholarWolf team (scholarwolf@library.unr.edu) with information about the object, including its full URL and the nature of your complaint."]},{"key":"dc:description.abstract","label":"Abstract","values":["A growing epidemic of pine tree deforestation in North America can largely be attributed to a particular pest, the Mountain Pine Beetle (MPB, Dendroctonus ponderosae). Coupling pheromone-coordinated mass attacks with an ability to resist the trees' natural defenses allows the beetles to overwhelm their targets and burrow beneath the tree bark to reproduce. MPB use cytochrome P450 enzymes to both detoxify toxic monoterpene compounds present in tree resin as well as synthesize the pheromones used during tree invasion, suggesting an evolutionary link between the P450 proteins involved in the two processes. This study attempts to contribute to the understanding of this evolutionary link by examining CYP6DH3, a P450 closely related to a known MPB resin detoxifier CYP6DH2. To do so, microsomal CYP6DH3 was expressed in Sf9 insect cells before being separated and incubated with various monoterpene substrates. GC screening revealed that no products could consistently be formed, which suggested an issue with the assay protocol. Various adjustments to the procedure have allowed the results of a previous study (Song et al., 2013) to be replicated, though further testing must be done to confirm whether these reaction conditions apply to CYP6DH3."]},{"key":"dc:format","label":"Dc Format","values":["PDF"]},{"key":"dc:title","label":"Title","values":["Resin Detoxification in Dendroctonus ponderosae: Functional Characterizations of Cytochrome P450 Proteins"]}]}],"canonical_facts":{"dc:contributor.advisor":["Tittiger, Claus R."],"dc:creator":["Bernardo, Joseph"],"dc:date.accessioned":["2017-01-24T23:09:18Z"],"dc:date.available":["2017-01-24T23:09:18Z"],"dc:date.issued":["2015"],"dc:description":["The University of Nevada, Reno Libraries will promptly respond to removal requests related to content that violates intellectual property laws, data protections, or has been uploaded without creator consent. Takedown notices should be directed to our ScholarWolf team (scholarwolf@library.unr.edu) with information about the object, including its full URL and the nature of your complaint."],"dc:description.abstract":["A growing epidemic of pine tree deforestation in North America can largely be attributed to a particular pest, the Mountain Pine Beetle (MPB, Dendroctonus ponderosae). Coupling pheromone-coordinated mass attacks with an ability to resist the trees' natural defenses allows the beetles to overwhelm their targets and burrow beneath the tree bark to reproduce. MPB use cytochrome P450 enzymes to both detoxify toxic monoterpene compounds present in tree resin as well as synthesize the pheromones used during tree invasion, suggesting an evolutionary link between the P450 proteins involved in the two processes. This study attempts to contribute to the understanding of this evolutionary link by examining CYP6DH3, a P450 closely related to a known MPB resin detoxifier CYP6DH2. To do so, microsomal CYP6DH3 was expressed in Sf9 insect cells before being separated and incubated with various monoterpene substrates. GC screening revealed that no products could consistently be formed, which suggested an issue with the assay protocol. Various adjustments to the procedure have allowed the results of a previous study (Song et al., 2013) to be replicated, though further testing must be done to confirm whether these reaction conditions apply to CYP6DH3."],"dc:format":["PDF"],"dc:identifier.uri":["http://hdl.handle.net/11714/641"],"dc:language":["English"],"dc:language.iso":["en_US"],"dc:rights":["In Copyright(All Rights Reserved)"],"dc:title":["Resin Detoxification in Dendroctonus ponderosae: Functional Characterizations of Cytochrome P450 Proteins"],"dc:type":["Thesis"],"thesis:degree_level":["Honors Thesis"],"thesis:degree_name":["Biochem and Molecular Biology"],"thesis:institution_name":["University of Nevada, Reno"]},"updated_at":"2026-07-27T21:46:08Z"}