Isolation and characterization of a synthetic gene and a genomic clone encoding acyl carrier protein from Escherichia coli

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 α-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.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Rawlings, Merriann

Contributors dc:contributor

• Cronan, John E.

• Biology, Molecular
• Biology, Microbiology

Statement dc:rights

• Copyright 1993 Rawlings, Merriann

Language dc:language

eng

Identifier

(UMI)AAI9411760
AAI9411760

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/20849