{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/20849"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/20849","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Isolation and characterization of a synthetic gene and a genomic clone encoding acyl carrier protein from Escherichia coli","abstract":"Acyl carrier protein (ACP) is a required cofactor for the synthesis and subsequent metabolism of fatty acids in Escherichia coli. Previous work has suggested that DNA segments encoding ACP were somehow toxic to E. coli. To investigate this possibility, a synthetic gene encoding ACP was assembled using a novel $lacZ\\ \\alpha$-complementation test. When provided with the sequences necessary for transcription and translation, the gene was expressed at high levels. Despite the already functional excess of ACP in wild-type cells its overexpression was indeed lethal, resulting in a decreased cellular growth rate and a precipitous drop in cell viability. One of the most obvious differences between ACP overproducing and wild-type strains was the accumulation of apo-ACP by the former cells. Normally apo-ACP is not detected in vivo. A genomic clone encoding ACP has also been isolated and sequenced. The ACP gene (called acpP) was located on the genetic map between fabF and fabD which encode two fatty acid biosynthetic enzymes, 3-ketoacyl-ACP synthase II and malonyl CoA-ACP transacylase, respectively. An open reading frame between acpP and fabD encodes a 26.5-kDa protein that has significant sequence identity ($>$40%) with a plant 3-ketoacyl-ACP reductase and thus is believed to encode the same enzyme in E. coli. This gene (called fabG) is cotranscribed with acpP. Thus, the gene encoding ACP is located within a cluster of fatty acid biosynthetic genes.","abstract_html":"Acyl carrier protein (ACP) is a required cofactor for the synthesis and subsequent metabolism of fatty acids in Escherichia coli. Previous work has suggested that DNA segments encoding ACP were somehow toxic to E. coli. To investigate this possibility, a synthetic gene encoding ACP was assembled using a novel <span class=\"etd-inline-math\">lacZ &alpha;</span>-complementation test. When provided with the sequences necessary for transcription and translation, the gene was expressed at high levels. Despite the already functional excess of ACP in wild-type cells its overexpression was indeed lethal, resulting in a decreased cellular growth rate and a precipitous drop in cell viability. One of the most obvious differences between ACP overproducing and wild-type strains was the accumulation of apo-ACP by the former cells. Normally apo-ACP is not detected in vivo. A genomic clone encoding ACP has also been isolated and sequenced. The ACP gene (called acpP) was located on the genetic map between fabF and fabD which encode two fatty acid biosynthetic enzymes, 3-ketoacyl-ACP synthase II and malonyl CoA-ACP transacylase, respectively. An open reading frame between acpP and fabD encodes a 26.5-kDa protein that has significant sequence identity ($&gt;$40%) with a plant 3-ketoacyl-ACP reductase and thus is believed to encode the same enzyme in E. coli. This gene (called fabG) is cotranscribed with acpP. Thus, the gene encoding ACP is located within a cluster of fatty acid biosynthetic genes.","abstract_has_math":true,"creators":["Rawlings, Merriann"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":["Cronan, John E."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T12:51:07Z","date_published":"2011-05-07T12:51:07Z","updated_at":"2026-07-22T22:25:16Z","subjects":["Biology, Molecular","Biology, Microbiology"],"languages":["eng"],"rights":["Copyright 1993 Rawlings, Merriann"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI9411760","AAI9411760"],"render_values":[{"text":"(UMI)AAI9411760","href":null,"code":true},{"text":"AAI9411760","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/20849","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Cronan, John E."]},{"key":"dc:creator","label":"Author","values":["Rawlings, Merriann"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T12:51:07Z","10000-01-01","1993"]},{"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 1993 Rawlings, Merriann"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI9411760","AAI9411760","http://hdl.handle.net/2142/20849"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Acyl carrier protein (ACP) is a required cofactor for the synthesis and subsequent metabolism of fatty acids in Escherichia coli. Previous work has suggested that DNA segments encoding ACP were somehow toxic to E. coli. To investigate this possibility, a synthetic gene encoding ACP was assembled using a novel $lacZ\\ \\alpha$-complementation test. When provided with the sequences necessary for transcription and translation, the gene was expressed at high levels. Despite the already functional excess of ACP in wild-type cells its overexpression was indeed lethal, resulting in a decreased cellular growth rate and a precipitous drop in cell viability. One of the most obvious differences between ACP overproducing and wild-type strains was the accumulation of apo-ACP by the former cells. Normally apo-ACP is not detected in vivo. A genomic clone encoding ACP has also been isolated and sequenced. The ACP gene (called acpP) was located on the genetic map between fabF and fabD which encode two fatty acid biosynthetic enzymes, 3-ketoacyl-ACP synthase II and malonyl CoA-ACP transacylase, respectively. An open reading frame between acpP and fabD encodes a 26.5-kDa protein that has significant sequence identity ($>$40%) with a plant 3-ketoacyl-ACP reductase and thus is believed to encode the same enzyme in E. coli. This gene (called fabG) is cotranscribed with acpP. Thus, the gene encoding ACP is located within a cluster of fatty acid biosynthetic genes.","Made available in DSpace on 2011-05-07T12:51:07Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9411760.pdf: 3217156 bytes, checksum: 4ffe98680c888f6a1deaad2e54ecf30c (MD5) Previous issue date: 1993","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:46:43Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:20:58-05:00 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":["Isolation and characterization of a synthetic gene and a genomic clone encoding acyl carrier protein from Escherichia coli"]}]}],"canonical_facts":{"dc:contributor":["Cronan, John E."],"dc:creator":["Rawlings, Merriann"],"dc:date":["2011-05-07T12:51:07Z","10000-01-01","1993"],"dc:description":["Acyl carrier protein (ACP) is a required cofactor for the synthesis and subsequent metabolism of fatty acids in Escherichia coli. Previous work has suggested that DNA segments encoding ACP were somehow toxic to E. coli. To investigate this possibility, a synthetic gene encoding ACP was assembled using a novel $lacZ\\ \\alpha$-complementation test. When provided with the sequences necessary for transcription and translation, the gene was expressed at high levels. Despite the already functional excess of ACP in wild-type cells its overexpression was indeed lethal, resulting in a decreased cellular growth rate and a precipitous drop in cell viability. One of the most obvious differences between ACP overproducing and wild-type strains was the accumulation of apo-ACP by the former cells. Normally apo-ACP is not detected in vivo. A genomic clone encoding ACP has also been isolated and sequenced. The ACP gene (called acpP) was located on the genetic map between fabF and fabD which encode two fatty acid biosynthetic enzymes, 3-ketoacyl-ACP synthase II and malonyl CoA-ACP transacylase, respectively. An open reading frame between acpP and fabD encodes a 26.5-kDa protein that has significant sequence identity ($>$40%) with a plant 3-ketoacyl-ACP reductase and thus is believed to encode the same enzyme in E. coli. This gene (called fabG) is cotranscribed with acpP. Thus, the gene encoding ACP is located within a cluster of fatty acid biosynthetic genes.","Made available in DSpace on 2011-05-07T12:51:07Z (GMT). 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