English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/40433
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Title

A new generation of vectors with increased induction ratios by overimposing a second regulatory level by attenuation

AuthorsRoyo, José Luis ; Manyani, Hamid; Cebolla, Ángel; Santero, Eduardo
KeywordsBacterial proteins
DNA-binding proteins
Gene expression regulation
Klebsiella oxytoca
Operon
Pseudomonas putida
RNA- binding
Trans-activators
Transcription factors
XylS protein
Pseudomonas putida
Antiterminator proteins
Issue Date31-Oct-2005
PublisherOxford University Press
CitationNucleic Acids Research 33(19): e169 (2005)
AbstractA major drawback of regulated gene expression from vectors bearing strong promoters is the associated high basal expression level. Simple regulatory systems have an intrinsic limitation in the range of induction, and attempts to mutate promoters to reduce basal expression usually result in concomitant reduction of induced levels. We have explored the possibility of reducing basal levels of gene expression while keeping induced levels intact by incorporating an additional regulatory circuit controlling a different step of the expression process. We have integrated the nasFEDCBA transcriptional attenuation system of Klebsiella oxytoca into a cascade expression circuit based on different regulatory elements of Pseudomonas putida, and also into a system based on the tac promoter, to expand their regulatory capacity. Basal expression from the promoters of these circuits was reduced by more than 10-fold by the nasF attenuator sequence while keeping the induced levels intact in the presence of the antiterminator protein, thus increasing the induction ratio by up to 1700-fold. In addition, using different combinations of regulatory elements and inducing conditions, we were able to obtain a broad range of expression levels. These vectors and the concept of their design will be very useful in regulating overproduction of heterologous proteins both at laboratory and industrial scales.
Description10 páginas, 3 figuras, 2 tablas.
Publisher version (URL)http://dx.doi.org/10.1093/nar/gni168
URIhttp://hdl.handle.net/10261/40433
DOI10.1093/nar/gni168
ISSN1362-1048
E-ISSN1362-4962
Appears in Collections:(CABD) Artículos
Files in This Item:
File Description SizeFormat 
Generation_2005.pdf304,15 kBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.