{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/18938"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/18938","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Mutational analysis of integrase and excisionase proteins in bacteriophage lambda site-specific recombination system","abstract":"Second-site reversion analyses were performed on 21 recombination defective Int mutants. A second-site revertant P243L/E218K and a pseudorevertant E175K were isolated and further characterized using genetic and biochemical methods. The results showed that the second-site substitution E218K selectively enhances Int core-binding affinity and acts as a global suppressor. In addition, our genetic data suggest that position 218 is part of or near the putative Int core-binding domain.","abstract_html":"Second-site reversion analyses were performed on 21 recombination defective Int mutants. A second-site revertant P243L/E218K and a pseudorevertant E175K were isolated and further characterized using genetic and biochemical methods. The results showed that the second-site substitution E218K selectively enhances Int core-binding affinity and acts as a global suppressor. In addition, our genetic data suggest that position 218 is part of or near the putative Int core-binding domain.","abstract_has_math":false,"creators":["Wu, Zhao"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":["Gardner, Jeffrey F.","Gumport, Richard I.","Voss, Edward W., Jr."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T11:51:59Z","date_published":"2011-05-07T11:51:59Z","updated_at":"2026-07-22T22:25:12Z","subjects":["Biology, Molecular","Biology, Microbiology"],"languages":["eng"],"rights":["Copyright 1995 Wu, Zhao"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9624540","(UMI)AAI9624540"],"render_values":[{"text":"AAI9624540","href":null,"code":true},{"text":"(UMI)AAI9624540","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/18938","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gardner, Jeffrey F.","Gumport, Richard I.","Voss, Edward W., Jr."]},{"key":"dc:creator","label":"Author","values":["Wu, Zhao"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T11:51:59Z","10000-01-01","1995"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"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":["Biology, Molecular","Biology, Microbiology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1995 Wu, Zhao"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9624540","(UMI)AAI9624540","http://hdl.handle.net/2142/18938"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Second-site reversion analyses were performed on 21 recombination defective Int mutants. A second-site revertant P243L/E218K and a pseudorevertant E175K were isolated and further characterized using genetic and biochemical methods. The results showed that the second-site substitution E218K selectively enhances Int core-binding affinity and acts as a global suppressor. In addition, our genetic data suggest that position 218 is part of or near the putative Int core-binding domain.","Saturation mutagenesis was performed on the region of the xis gene that encodes carboxyl region of the Xis protein. Forty-five new C-terminal region mutants were isolated and their abilities to promote excision, to bind the DNA and to interact with Int and FIS were characterized in vivo and in vitro. In addition, the secondary structure of this region was predicted by computer modeling schemes. A model for the secondary structures of carboxyl region of Xis protein was developed by combining the mutagenesis information with computer-based analytic techniques. According to the model, residues between 57 to 65 are located in an $\\alpha$-helix structure. The functional study of mutants within the helix suggests that one side of the helix (residues 57, 60, 63 and 64) interacts with Int. In addition, the identification of structurally stable mutants of Xis may be useful in studying its degradation pathway and in supplying protein for crystallographic studies.","Made available in DSpace on 2011-05-07T11:51:59Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9624540.pdf: 5869368 bytes, checksum: d9b46729729a489ce90abcfaf05010c0 (MD5) Previous issue date: 1995","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:33:33Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:12:26-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"]},{"key":"dc:title","label":"Title","values":["Mutational analysis of integrase and excisionase proteins in bacteriophage lambda site-specific recombination system"]}]}],"canonical_facts":{"dc:contributor":["Gardner, Jeffrey F.","Gumport, Richard I.","Voss, Edward W., Jr."],"dc:creator":["Wu, Zhao"],"dc:date":["2011-05-07T11:51:59Z","10000-01-01","1995"],"dc:description":["Second-site reversion analyses were performed on 21 recombination defective Int mutants. A second-site revertant P243L/E218K and a pseudorevertant E175K were isolated and further characterized using genetic and biochemical methods. The results showed that the second-site substitution E218K selectively enhances Int core-binding affinity and acts as a global suppressor. In addition, our genetic data suggest that position 218 is part of or near the putative Int core-binding domain.","Saturation mutagenesis was performed on the region of the xis gene that encodes carboxyl region of the Xis protein. Forty-five new C-terminal region mutants were isolated and their abilities to promote excision, to bind the DNA and to interact with Int and FIS were characterized in vivo and in vitro. In addition, the secondary structure of this region was predicted by computer modeling schemes. A model for the secondary structures of carboxyl region of Xis protein was developed by combining the mutagenesis information with computer-based analytic techniques. According to the model, residues between 57 to 65 are located in an $\\alpha$-helix structure. The functional study of mutants within the helix suggests that one side of the helix (residues 57, 60, 63 and 64) interacts with Int. In addition, the identification of structurally stable mutants of Xis may be useful in studying its degradation pathway and in supplying protein for crystallographic studies.","Made available in DSpace on 2011-05-07T11:51:59Z (GMT). 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