{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/102515"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/102515","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Development of in vivo ppGpp reporters in escherichia coli","abstract":"ppGpp is a small molecule that works as a global master regulator of the E. coli physiology, most notably during the stringent response. Systems biology studies have emphasized its role in understanding the host response to the burden of introduced synthetic genetic circuits. Traditionally, researchers rely on in vitro methods to measure the intracellular ppGpp levels. In vivo ppGpp reporters, however, would allow the possibility of closely monitoring the ppGpp levels during different processes, especially the host response given the functioning of synthetic circuits with relatively fast dynamics. In this study, a series of in vivo ppGpp reporters were developed in E. coli by placing a fluorescent protein under the control of promoters from genes known to be controlled by ppGpp levels. Reporter performance was tested by monitoring the fluorescence level with microscope during different stages of growth, induced stringent response, and different growth conditions. The four positive ppGpp reporters showed strong positive correlation with ppGpp levels, including two with particularly high sensitivity and signal intensity. The testing process also shed new light on the dynamics of ppGpp production during batch culture growth and under nutrient limitation. With further development, these in vivo ppGpp reporters promise to be very useful when studying the dynamics of host resource partitioning.","abstract_html":"ppGpp is a small molecule that works as a global master regulator of the E. coli physiology, most notably during the stringent response. Systems biology studies have emphasized its role in understanding the host response to the burden of introduced synthetic genetic circuits. Traditionally, researchers rely on in vitro methods to measure the intracellular ppGpp levels. In vivo ppGpp reporters, however, would allow the possibility of closely monitoring the ppGpp levels during different processes, especially the host response given the functioning of synthetic circuits with relatively fast dynamics. In this study, a series of in vivo ppGpp reporters were developed in E. coli by placing a fluorescent protein under the control of promoters from genes known to be controlled by ppGpp levels. Reporter performance was tested by monitoring the fluorescence level with microscope during different stages of growth, induced stringent response, and different growth conditions. The four positive ppGpp reporters showed strong positive correlation with ppGpp levels, including two with particularly high sensitivity and signal intensity. The testing process also shed new light on the dynamics of ppGpp production during batch culture growth and under nutrient limitation. With further development, these in vivo ppGpp reporters promise to be very useful when studying the dynamics of host resource partitioning.","abstract_has_math":false,"creators":["Li, Jinbei Walden"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"M.S.","degree_level":"Thesis","degree_discipline":"Bioengineering","degree_department":null,"school":null,"contributors":["Lu, Ting"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-02-06T19:36:43Z","date_published":"2019-02-06T19:36:43Z","updated_at":"2026-07-22T22:24:42Z","subjects":["ESCHERICHIA COLI, ppGpp"],"languages":["en"],"rights":["Copyright 2018 Jinbei Li"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/102515","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Lu, Ting"]},{"key":"dc:creator","label":"Author","values":["Li, Jinbei Walden"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-02-06T19:36:43Z","2018-12-14","2018-12"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Bioengineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["ESCHERICHIA COLI, ppGpp"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2018 Jinbei Li"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/102515"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["ppGpp is a small molecule that works as a global master regulator of the E. coli physiology, most notably during the stringent response. Systems biology studies have emphasized its role in understanding the host response to the burden of introduced synthetic genetic circuits. Traditionally, researchers rely on in vitro methods to measure the intracellular ppGpp levels. In vivo ppGpp reporters, however, would allow the possibility of closely monitoring the ppGpp levels during different processes, especially the host response given the functioning of synthetic circuits with relatively fast dynamics. In this study, a series of in vivo ppGpp reporters were developed in E. coli by placing a fluorescent protein under the control of promoters from genes known to be controlled by ppGpp levels. Reporter performance was tested by monitoring the fluorescence level with microscope during different stages of growth, induced stringent response, and different growth conditions. The four positive ppGpp reporters showed strong positive correlation with ppGpp levels, including two with particularly high sensitivity and signal intensity. The testing process also shed new light on the dynamics of ppGpp production during batch culture growth and under nutrient limitation. With further development, these in vivo ppGpp reporters promise to be very useful when studying the dynamics of host resource partitioning.","Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2019-02-05 without embargo terms","The student, Jinbei Li, accepted the attached license on 2018-12-14 at 12:19.","The student, Jinbei Li, submitted this Thesis for approval on 2018-12-14 at 12:20.","This Thesis was approved for publication on 2018-12-14 at 12:48.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13295 on 2019-02-05 at 11:16:06","Made available in DSpace on 2019-02-06T19:36:43Z (GMT). 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In vivo ppGpp reporters, however, would allow the possibility of closely monitoring the ppGpp levels during different processes, especially the host response given the functioning of synthetic circuits with relatively fast dynamics. In this study, a series of in vivo ppGpp reporters were developed in E. coli by placing a fluorescent protein under the control of promoters from genes known to be controlled by ppGpp levels. Reporter performance was tested by monitoring the fluorescence level with microscope during different stages of growth, induced stringent response, and different growth conditions. The four positive ppGpp reporters showed strong positive correlation with ppGpp levels, including two with particularly high sensitivity and signal intensity. The testing process also shed new light on the dynamics of ppGpp production during batch culture growth and under nutrient limitation. With further development, these in vivo ppGpp reporters promise to be very useful when studying the dynamics of host resource partitioning.","Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2019-02-05 without embargo terms","The student, Jinbei Li, accepted the attached license on 2018-12-14 at 12:19.","The student, Jinbei Li, submitted this Thesis for approval on 2018-12-14 at 12:20.","This Thesis was approved for publication on 2018-12-14 at 12:48.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13295 on 2019-02-05 at 11:16:06","Made available in DSpace on 2019-02-06T19:36:43Z (GMT). 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