{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/21092"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/21092","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Biochemical and genetic analysis of nucleic acid-protein interactions","abstract":"The integration host factor (IHF) of Escherichia coli is a small, sequence-specific DNA-binding protein. The specific and nonspecific DNA binding constants were determined by gel mobility shift assays. The binding constant of IHF for the H$\\sp\\prime$ site in $\\lambda$ attP is 6.8 $\\times$ 10$\\sp8$ M$\\sp{-1}$ and the nonspecific binding constant is 5.8 $\\times$ 10$\\sp5$ M$\\sp{-1}.$ Therefore, the selectivity of IHF binding is about a thousand-fold higher for a specific site over random sequences. To study the molecular determinants specifying IHF binding, a series of 41 oligonucleotides were synthesized containing adenine analogues that modified the surfaces of the major and minor grooves of DNA. The binding constants determined for these analogue-containing H$\\sp\\prime$ sites revealed that the specific interaction of IHF with its H$\\sp\\prime$ site involves interactions with both minor and major grooves of the DNA.","abstract_html":"The integration host factor (IHF) of Escherichia coli is a small, sequence-specific DNA-binding protein. The specific and nonspecific DNA binding constants were determined by gel mobility shift assays. The binding constant of IHF for the H$\\sp\\prime$ site in $\\lambda$ attP is 6.8 $\\times$ 10$\\sp8$ M$\\sp{-1}$ and the nonspecific binding constant is 5.8 $\\times$ 10$\\sp5$ M$\\sp{-1}.$ Therefore, the selectivity of IHF binding is about a thousand-fold higher for a specific site over random sequences. To study the molecular determinants specifying IHF binding, a series of 41 oligonucleotides were synthesized containing adenine analogues that modified the surfaces of the major and minor grooves of DNA. The binding constants determined for these analogue-containing H$\\sp\\prime$ sites revealed that the specific interaction of IHF with its H$\\sp\\prime$ site involves interactions with both minor and major grooves of the DNA.","abstract_has_math":true,"creators":["Wang, Siqun"],"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."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T12:58:08Z","date_published":"2011-05-07T12:58:08Z","updated_at":"2026-07-22T22:25:17Z","subjects":["Biology, Molecular","Biology, Genetics","Biology, Microbiology"],"languages":["eng"],"rights":["Copyright 1995 Wang, Siqun"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9624530","(UMI)AAI9624530"],"render_values":[{"text":"AAI9624530","href":null,"code":true},{"text":"(UMI)AAI9624530","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/21092","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Gardner, Jeffrey F."]},{"key":"dc:creator","label":"Author","values":["Wang, Siqun"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T12:58:08Z","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, Genetics","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 Wang, Siqun"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9624530","(UMI)AAI9624530","http://hdl.handle.net/2142/21092"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The integration host factor (IHF) of Escherichia coli is a small, sequence-specific DNA-binding protein. The specific and nonspecific DNA binding constants were determined by gel mobility shift assays. The binding constant of IHF for the H$\\sp\\prime$ site in $\\lambda$ attP is 6.8 $\\times$ 10$\\sp8$ M$\\sp{-1}$ and the nonspecific binding constant is 5.8 $\\times$ 10$\\sp5$ M$\\sp{-1}.$ Therefore, the selectivity of IHF binding is about a thousand-fold higher for a specific site over random sequences. To study the molecular determinants specifying IHF binding, a series of 41 oligonucleotides were synthesized containing adenine analogues that modified the surfaces of the major and minor grooves of DNA. The binding constants determined for these analogue-containing H$\\sp\\prime$ sites revealed that the specific interaction of IHF with its H$\\sp\\prime$ site involves interactions with both minor and major grooves of the DNA.","The coat protein of coliphage R17 is a small, sequence-specific RNA-binding protein. The ribophage system was used to characterize the specific interactions between the coat protein and its RNA-binding site. Studies with the RNA-binding site suggest that the translational repression conferred by this hairpin depends on its interactions with both the R17 coat protein and the ribosome. In addition, a total of eight R17 coat protein mutants were isolated with altered, expanded RNA-binding activities in vivo. They contain substitutions at the N-terminus and in the middle of the coat protein. Molecular characterizations of these coat protein mutants suggest that there are two classes of mutants. Possible mechanisms of the in vivo suppression of phage variants by these mutant coat proteins are discussed.","Made available in DSpace on 2011-05-07T12:58:08Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9624530.pdf: 4012896 bytes, checksum: b1e378b542a0292358deb6aabac1eefb (MD5) Previous issue date: 1995","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:48:25Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:21:55-05:00 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":["Biochemical and genetic analysis of nucleic acid-protein interactions"]}]}],"canonical_facts":{"dc:contributor":["Gardner, Jeffrey F."],"dc:creator":["Wang, Siqun"],"dc:date":["2011-05-07T12:58:08Z","10000-01-01","1995"],"dc:description":["The integration host factor (IHF) of Escherichia coli is a small, sequence-specific DNA-binding protein. The specific and nonspecific DNA binding constants were determined by gel mobility shift assays. The binding constant of IHF for the H$\\sp\\prime$ site in $\\lambda$ attP is 6.8 $\\times$ 10$\\sp8$ M$\\sp{-1}$ and the nonspecific binding constant is 5.8 $\\times$ 10$\\sp5$ M$\\sp{-1}.$ Therefore, the selectivity of IHF binding is about a thousand-fold higher for a specific site over random sequences. To study the molecular determinants specifying IHF binding, a series of 41 oligonucleotides were synthesized containing adenine analogues that modified the surfaces of the major and minor grooves of DNA. The binding constants determined for these analogue-containing H$\\sp\\prime$ sites revealed that the specific interaction of IHF with its H$\\sp\\prime$ site involves interactions with both minor and major grooves of the DNA.","The coat protein of coliphage R17 is a small, sequence-specific RNA-binding protein. The ribophage system was used to characterize the specific interactions between the coat protein and its RNA-binding site. Studies with the RNA-binding site suggest that the translational repression conferred by this hairpin depends on its interactions with both the R17 coat protein and the ribosome. In addition, a total of eight R17 coat protein mutants were isolated with altered, expanded RNA-binding activities in vivo. They contain substitutions at the N-terminus and in the middle of the coat protein. Molecular characterizations of these coat protein mutants suggest that there are two classes of mutants. Possible mechanisms of the in vivo suppression of phage variants by these mutant coat proteins are discussed.","Made available in DSpace on 2011-05-07T12:58:08Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9624530.pdf: 4012896 bytes, checksum: b1e378b542a0292358deb6aabac1eefb (MD5) Previous issue date: 1995","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:48:25Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:21:55-05:00 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"],"dc:identifier":["AAI9624530","(UMI)AAI9624530","http://hdl.handle.net/2142/21092"],"dc:language":["eng"],"dc:rights":["Copyright 1995 Wang, Siqun"],"dc:subject":["Biology, Molecular","Biology, Genetics","Biology, Microbiology"],"dc:title":["Biochemical and genetic analysis of nucleic acid-protein interactions"],"dc:type":["text"],"thesis:degree_discipline":["Microbiology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:17Z"}