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oai:www.ideals.illinois.edu:2142/20849 2023-07-10 col_2142_5131 col_2142_16338 com_2142_5130 com_2142_16337 com_2142_14793 com_2142_8903

(UMI)AAI9411760 AAI9411760 Cronan, John E. Rawlings, Merriann 2011-05-07T12:51:07Z 2011-05-07T12:51:07Z 10000-01-01 1993 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 http://hdl.handle.net/2142/20849 eng Copyright 1993 Rawlings, Merriann Biology, Molecular Biology, Microbiology Isolation and characterization of a synthetic gene and a genomic clone encoding acyl carrier protein from Escherichia coli text Microbiology Microbiology University of Illinois at Urbana-Champaign Dissertation Ph.D.