Document Detail


Plasmid stability of recombinant Pseudomonas sp. B13 FR1 pFRC20P in continuous culture.
MedLine Citation:
PMID:  10099179     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Plasmid stability of recombinant Pseudomonas sp. B13 FR1 pFRC20P, a strain capable of mineralizing 3- and 4-chlorobenzoate and 4-methylbenzoate, was investigated in continuous culture. The hybrid cosmid pFRC20P enables the strain to mineralize 4-methylbenzoate. Rapid plasmid loss was observed under nonselective conditions using 3-chlorobenzoate as the substrate. Plasmid stability decreased with increasing dilution rate. Despite the growth advantage of the generated plasmid free cells a total depletion of plasmid bearing cells was not observed. After approximately 50 generations the fraction of plasmid bearing cells reached a constant level of 10%, which was stably maintained during the next 25 generations. Cells from this stage were used to inoculate a new culture that resulted in a stable level of 50% plasmid bearing cells. By a temporary substrate change to selective conditions (4-methylbenzoate), this level could be further increased to 70%. Literature models on plasmid stability could not be applied to describe the experimental data. Therefore, a new but unstructured model was developed to describe the experimental results. The model is based on the existence of three subpopulations: a plasmid free one, an original plasmid bearing one with a growth disadvantage compared to plasmid free cells, and a second plasmid bearing subpopulation with increased stability that is generated from the original one and has a growth rate comparable to the plasmid free cells.
Authors:
C Hempel; R W Erb; W Deckwer; V Hecht
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  57     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  1998 Jan 
Date Detail:
Created Date:  1999-05-07     Completed Date:  1999-05-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  62-70     Citation Subset:  IM    
Copyright Information:
Copyright 1998 John Wiley & Sons, Inc.
Affiliation:
Biochemical Engineering Division, GBF-Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, 38124 Braunschweig, Germany.
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MeSH Terms
Descriptor/Qualifier:
Benzoic Acids / metabolism
Biodegradation, Environmental
Bioreactors
Biotechnology
Chlorobenzoates / metabolism
Models, Biological
Plasmids / genetics*
Pseudomonas / genetics*,  metabolism
Recombination, Genetic
Chemical
Reg. No./Substance:
0/Benzoic Acids; 0/Chlorobenzoates; 535-80-8/3-chlorobenzoic acid; 74-11-3/4-chlorobenzoic acid; 99-94-5/4-toluic acid

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