Document Detail


An experimental and theoretical study of the inhibition of Escherichia coli lac operon gene expression by antigene oligonucleotides.
MedLine Citation:
PMID:  11400095     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previously, we have developed a genetically structured mathematical model to describe the inhibition of Escherichia coli lac operon gene expression by antigene oligos. Our model predicted that antigene oligos targeted to the operator region of the lac operon would have a significant inhibitory effect on beta-galactosidase production. In this investigation, the E. coli lac operon gene expression in the presence of antigene oligos was studied experimentally. A 21-mer oligo, which was designed to form a triplex with the operator, was found to be able to specifically inhibit beta-galactosidase production in a dose-dependent manner. In contrast to the 21-mer triplex-forming oligonucleotide (TFO), several control oligos showed no inhibitory effect. The ineffectiveness of the various control oligos, along with the fact that the 21-mer oligo has no homology sequence with lacZYA, and no mRNA is transcribed from the operator, suggests that the 21-mer oligo inhibits target gene expression by an antigene mechanism. To simulate the kinetics of lac operon gene expression in the presence of antigene oligos, a genetically structured kinetic model, which includes transport of oligo into the cell, growth of bacteria cells, and lac operon gene expression, was developed. Predictions of the kinetic model fit the experimental data quite well after adjustment of the value of the oligonucleotide transport rate constant (9.0 x 10(-)(3) min(-)(1)) and oligo binding affinity constant (1.05 x 10(6) M(-)(1)). Our values for these two adjusted parameters are in the range of reported literature values.
Authors:
B Cheng; R L Fournier; P A Relue; J Schisler
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  74     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-06-11     Completed Date:  2001-10-04     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  220-9     Citation Subset:  IM    
Copyright Information:
Copyright 2001 John Wiley & Sons, Inc.
Affiliation:
Department of Bioengineering, University of Toledo, Toledo, Ohio 43606, USA.
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MeSH Terms
Descriptor/Qualifier:
Biological Transport
Cell Division / drug effects,  genetics
Computer Simulation
Cyclic AMP / metabolism
Escherichia coli / drug effects,  genetics*,  metabolism
Gene Expression Regulation, Bacterial / drug effects
Isopropyl Thiogalactoside / pharmacokinetics
Kinetics
Models, Theoretical*
Oligonucleotides, Antisense / pharmacology*
Operon*
beta-Galactosidase / genetics
Chemical
Reg. No./Substance:
0/Oligonucleotides, Antisense; 367-93-1/Isopropyl Thiogalactoside; 60-92-4/Cyclic AMP; EC 3.2.1.23/beta-Galactosidase

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