{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/71187"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/71187","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Molecular Genetic Analysis of Rhodobacter Sphaeroides Light-Harvesting Complexes","abstract":"The genes for the Rhodobacter sphaeroides light-harvesting, B875-$\\beta$ and B875-$\\alpha$ polypeptides (pufB and pufA) are closely linked to the genes for the reaction center-L and reaction center-M polypeptides (pufL and pufM) on what has been termed the puf operon (gene order pufB,A,L,M). The DNA sequence of pufB and pufA from wild-type R. sphaeroides 2.4.1 was determined. The relative levels of the B875-$\\beta$ and B875-$\\alpha$ and the reaction center-L and reaction center-M polypeptides synthesized in a homologous cell-free transcription-translation system were compared with those found in vivo. Analysis of the gene products produced in vitro using plasmids containing deletions upstream of the pufB structural gene identified a region of DNA required for expression of the B875-$\\beta$ and B875-$\\alpha$ polypeptides. These results support the hypothesis that the mapped 5$\\sp\\prime$ termini of the large and small puf operon transcripts represent transcription initiation sites.","abstract_html":"The genes for the Rhodobacter sphaeroides light-harvesting, B875-<span class=\"etd-inline-math\">&beta;</span> and B875-<span class=\"etd-inline-math\">&alpha;</span> polypeptides (pufB and pufA) are closely linked to the genes for the reaction center-L and reaction center-M polypeptides (pufL and pufM) on what has been termed the puf operon (gene order pufB,A,L,M). The DNA sequence of pufB and pufA from wild-type R. sphaeroides 2.4.1 was determined. The relative levels of the B875-<span class=\"etd-inline-math\">&beta;</span> and B875-<span class=\"etd-inline-math\">&alpha;</span> and the reaction center-L and reaction center-M polypeptides synthesized in a homologous cell-free transcription-translation system were compared with those found in vivo. Analysis of the gene products produced in vitro using plasmids containing deletions upstream of the pufB structural gene identified a region of DNA required for expression of the B875-<span class=\"etd-inline-math\">&beta;</span> and B875-<span class=\"etd-inline-math\">&alpha;</span> polypeptides. These results support the hypothesis that the mapped 5$\\sp\\prime$ termini of the large and small puf operon transcripts represent transcription initiation sites.","abstract_has_math":true,"creators":["Kiley, Patricia Jeannette"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":["Kaplan, Samuel"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-12-16T06:13:00Z","date_published":"2014-12-16T06:13:00Z","updated_at":"2026-07-22T22:26:04Z","subjects":["Biology, Microbiology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI8803085"],"render_values":[{"text":"(UMI)AAI8803085","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/71187","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Kaplan, Samuel"]},{"key":"dc:creator","label":"Author","values":["Kiley, Patricia Jeannette"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2014-12-16T06:13:00Z","10000-01-01","1987"]},{"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, Microbiology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/71187","(UMI)AAI8803085"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The genes for the Rhodobacter sphaeroides light-harvesting, B875-$\\beta$ and B875-$\\alpha$ polypeptides (pufB and pufA) are closely linked to the genes for the reaction center-L and reaction center-M polypeptides (pufL and pufM) on what has been termed the puf operon (gene order pufB,A,L,M). The DNA sequence of pufB and pufA from wild-type R. sphaeroides 2.4.1 was determined. The relative levels of the B875-$\\beta$ and B875-$\\alpha$ and the reaction center-L and reaction center-M polypeptides synthesized in a homologous cell-free transcription-translation system were compared with those found in vivo. Analysis of the gene products produced in vitro using plasmids containing deletions upstream of the pufB structural gene identified a region of DNA required for expression of the B875-$\\beta$ and B875-$\\alpha$ polypeptides. These results support the hypothesis that the mapped 5$\\sp\\prime$ termini of the large and small puf operon transcripts represent transcription initiation sites.","Two deoxyoligonucleotide probes were synthesized in accordance with the available amino acid sequence of the B800-850-$\\beta$ polypeptide from Rhodobacter sphaeroides and were used to isolate a 2.6 kilobase PstI fragment from R. sphaeroides 2.4.1 genomic DNA. Identification of the B800-850-$\\beta$ and B800-850-$\\alpha$ structural genes, pucB and pucA, was confirmed by DNA sequencing. Northern blot analysis, using a pucB,A-specific restriction endonuclease fragment as a probe, revealed a single puc-operon-specific, highly stable, transcript of approximately 640 bases present in photosynthetically growing cells. In vitro transcription-translation analysis of the puc operon revealed maximum synthesis of the puc operon gene products was achieved using the 2.6 kb PstI fragment as template although a 537 bp XmaIII fragment was sufficient to direct the synthesis of pucB and a pucA fusion product.","Mutants deficient in either B800-850 or B875 spectral activity were characterized. The B800-850$\\sp-$ mutant also lacked visibly absorbing carotenoids. Moreover, B800-850-$\\alpha$ and B800-850-$\\beta$ polypeptides could not be detected in the photosynthetic membranes derived from the B800-850$\\sp-$ mutant. Reduced levels of the B875-$\\alpha$ polypeptide were present in the photosynthetic membranes from the B875$\\sp-$ mutant, although there was no detectable absorption at 875 nm. Both mutants responded similarly to the wild type in their response to changes in incident light-intensity. However, the magnitude of derepression for B800-850 synthesis was greater in the B875$\\sp-$ mutant than the wild-type when cells grown at equivalent light intensities were examined.","Made available in DSpace on 2014-12-16T06:13:00Z (GMT). No. of bitstreams: 1 8803085.pdf: 5673242 bytes, checksum: f40088c96d7ff5b777a355011a9b7187 (MD5) Previous issue date: 1987","Embargo set by: Seth Robbins for item 71353 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","180 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1987."]},{"key":"dc:title","label":"Title","values":["Molecular Genetic Analysis of Rhodobacter Sphaeroides Light-Harvesting Complexes"]}]}],"canonical_facts":{"dc:contributor":["Kaplan, Samuel"],"dc:creator":["Kiley, Patricia Jeannette"],"dc:date":["2014-12-16T06:13:00Z","10000-01-01","1987"],"dc:description":["The genes for the Rhodobacter sphaeroides light-harvesting, B875-$\\beta$ and B875-$\\alpha$ polypeptides (pufB and pufA) are closely linked to the genes for the reaction center-L and reaction center-M polypeptides (pufL and pufM) on what has been termed the puf operon (gene order pufB,A,L,M). The DNA sequence of pufB and pufA from wild-type R. sphaeroides 2.4.1 was determined. The relative levels of the B875-$\\beta$ and B875-$\\alpha$ and the reaction center-L and reaction center-M polypeptides synthesized in a homologous cell-free transcription-translation system were compared with those found in vivo. Analysis of the gene products produced in vitro using plasmids containing deletions upstream of the pufB structural gene identified a region of DNA required for expression of the B875-$\\beta$ and B875-$\\alpha$ polypeptides. These results support the hypothesis that the mapped 5$\\sp\\prime$ termini of the large and small puf operon transcripts represent transcription initiation sites.","Two deoxyoligonucleotide probes were synthesized in accordance with the available amino acid sequence of the B800-850-$\\beta$ polypeptide from Rhodobacter sphaeroides and were used to isolate a 2.6 kilobase PstI fragment from R. sphaeroides 2.4.1 genomic DNA. Identification of the B800-850-$\\beta$ and B800-850-$\\alpha$ structural genes, pucB and pucA, was confirmed by DNA sequencing. Northern blot analysis, using a pucB,A-specific restriction endonuclease fragment as a probe, revealed a single puc-operon-specific, highly stable, transcript of approximately 640 bases present in photosynthetically growing cells. In vitro transcription-translation analysis of the puc operon revealed maximum synthesis of the puc operon gene products was achieved using the 2.6 kb PstI fragment as template although a 537 bp XmaIII fragment was sufficient to direct the synthesis of pucB and a pucA fusion product.","Mutants deficient in either B800-850 or B875 spectral activity were characterized. The B800-850$\\sp-$ mutant also lacked visibly absorbing carotenoids. Moreover, B800-850-$\\alpha$ and B800-850-$\\beta$ polypeptides could not be detected in the photosynthetic membranes derived from the B800-850$\\sp-$ mutant. Reduced levels of the B875-$\\alpha$ polypeptide were present in the photosynthetic membranes from the B875$\\sp-$ mutant, although there was no detectable absorption at 875 nm. Both mutants responded similarly to the wild type in their response to changes in incident light-intensity. However, the magnitude of derepression for B800-850 synthesis was greater in the B875$\\sp-$ mutant than the wild-type when cells grown at equivalent light intensities were examined.","Made available in DSpace on 2014-12-16T06:13:00Z (GMT). No. of bitstreams: 1 8803085.pdf: 5673242 bytes, checksum: f40088c96d7ff5b777a355011a9b7187 (MD5) Previous issue date: 1987","Embargo set by: Seth Robbins for item 71353 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","180 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1987."],"dc:identifier":["http://hdl.handle.net/2142/71187","(UMI)AAI8803085"],"dc:subject":["Biology, Microbiology"],"dc:title":["Molecular Genetic Analysis of Rhodobacter Sphaeroides Light-Harvesting Complexes"],"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":"2026-07-22T22:26:04Z"}