Abstract:
BACKGROUND:Messenger RNA decay is an important mechanism for controlling gene expression in all organisms. The rate of the mRNA degradation directly affects the steady state concentration of mRNAs and therefore influences the protein synthesis. RNaseE has a key importance for the general mRNA decay in E. coli. While RNaseE initiates the degradation of most mRNAs in E. coli, it is likely that the enzyme is also responsible for the degradation of recombinant RNAs. As RNaseE is essential for cell viability and knockout mutants cannot be cultured, we investigated the possibility for a down-regulation of the intracellular level of RNaseE by antisense RNAs. During this study, an antisense RNA based approach could be established which revealed a strong reduction of the intracellular level of RNaseE in E. coli. RESULTS:Despite the autoregulation of rne mRNA by its gene product, significant antisense downregulation of RNaseE is possible. The expression of antisense RNAs did not effect the cell growth negatively. The amount of antisense RNA was monitored quantitatively by a fluorescence based sandwich hybridisation assay. Induction by anhydrotetracycline was followed by a 25-fold increase of the detectable antisense RNA molecules per cell. The antisense RNA level was maintained above 400 molecules per cell until the stationary phase, which caused the level of expressed antisense RNAs to decrease markedly. Western blot experiments revealed the strongest reduction in the RNaseE protein level 90 min after antisense RNA induction. The cellular level of RNaseE could be decreased to 35% of the wild type level. When the growth entered the stationary phase, the RNaseE level was maintained still at 50 to 60% of the wild type level. CONCLUSION:In difference to eukaryotic cells, where the RNAi technology is widely used, this technology is rather unexplored in bacteria, although different natural systems use antisense RNA-based silencing, and a few studies have earlier indicated the potential of this technology also in prokaryotes. Our results show that even complicated self-regulatory systems such as RNaseE may be controlled by antisense RNA technology, indicating that systems based on antisense RNA expression may have a potential for controlling detrimental factors with plasmid-based constructs in arbitrary strains while maintaining their beneficial characteristics. The study also proved that the RNA sandwich hybridisation technique is directly applicable to quantify small RNA molecules in crude cell extracts, which may have a broader application potential as a monitoring tool in RNA inhibition applications.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Kemmer C,Neubauer Pdoi
10.1186/1475-2859-5-38subject
Has Abstractpub_date
2006-12-12 00:00:00pages
38issn
1475-2859pii
1475-2859-5-38journal_volume
5pub_type
杂志文章abstract:BACKGROUND:p-Hydroxycinnamic acid (pHCA) is an aromatic compound that serves as a starting material for the production of many commercially valuable chemicals, such as fragrances and pharmaceuticals, and is also used in the synthesis of thermostable polymers. However, chemical synthesis of pHCA is both costly and harmf...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-45
更新日期:2013-05-07 00:00:00
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-44
更新日期:2013-05-04 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/1475-2859-13-S1-S10
更新日期:2014-08-29 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0984-x
更新日期:2018-09-01 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-30
更新日期:2007-09-19 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1198-6
更新日期:2019-09-04 00:00:00
abstract:BACKGROUND:E. coli is a robust host for various genetic manipulations and has been used commonly for bioconversion of hexose and pentose sugars into valuable products. One of the products that E. coli make under fermentative condition is ethanol. However, availability of limited reducing equivalence and generation of c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-145
更新日期:2012-11-04 00:00:00
abstract:BACKGROUND:Mannosylglycerate (MG) is one of the most widespread compatible solutes among marine microorganisms adapted to hot environments. This ionic solute holds excellent ability to protect proteins against thermal denaturation, hence a large number of biotechnological and clinical applications have been put forward...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-1023-7
更新日期:2018-11-16 00:00:00
abstract:BACKGROUND:In Saccharomyces cerevisiae, alpha-glucosidase (maltase) is a key enzyme in maltose metabolism. In addition, the overexpression of the alpha-glucosidase-encoding gene MAL62 has been shown to increase the freezing tolerance of yeast in lean dough. However, its cryoprotection mechanism is still not clear. RES...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01454-6
更新日期:2020-10-19 00:00:00
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pub_type: 杂志文章
doi:10.1186/1475-2859-10-67
更新日期:2011-08-11 00:00:00
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pub_type: 杂志文章
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更新日期:2015-11-26 00:00:00
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pub_type: 杂志文章
doi:10.1186/s12934-014-0095-2
更新日期:2014-08-12 00:00:00
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pub_type: 杂志文章
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更新日期:2018-02-22 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-30
更新日期:2010-05-11 00:00:00
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pub_type: 杂志文章
doi:10.1186/1475-2859-11-138
更新日期:2012-10-24 00:00:00
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pub_type: 杂志文章
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更新日期:2018-01-13 00:00:00
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更新日期:2019-03-28 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2019-09-21 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2017-05-31 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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