Construction of in vitro transcription system for Corynebacterium glutamicum and its use in the recognition of promoters of different classes

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F12%3A00383121" target="_blank" >RIV/61388971:_____/12:00383121 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1007/s00253-012-4336-1" target="_blank" >http://dx.doi.org/10.1007/s00253-012-4336-1</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00253-012-4336-1" target="_blank" >10.1007/s00253-012-4336-1</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Construction of in vitro transcription system for Corynebacterium glutamicum and its use in the recognition of promoters of different classes

Popis výsledku v původním jazyce

To facilitate transcription studies in Corynebacterium glutamicum, we have developed an in vitro transcription system for this bacterium used as an industrial producer of amino acids and a model organism for actinobacteria. This system consists of C. glutamicum RNA polymerase (RNAP) core (alpha 2, beta, beta'), a sigma factor and a promoter-carrying DNA template, that is specifically recognized by the RNAP holoenzyme formed. The RNAP core was purified from the C. glutamicum strain with the modified rpoCgene, which produced His-tagged beta' subunit. The C. glutamicum sigA and sigH genes were cloned and overexpressed using the Escherichia coli plasmid vector, and the sigma subunits sigma(A) and sigma(H) were purified by affinity chromatography. Using the reconstituted C. glutamicum holo-RNAPs, recognition of the sigma(A)- and sigma(H)-dependent promoters and synthesis of the specific transcripts was demonstrated. The developed in vitro transcription system is a novel tool that can be us

Název v anglickém jazyce

Construction of in vitro transcription system for Corynebacterium glutamicum and its use in the recognition of promoters of different classes

Popis výsledku anglicky

To facilitate transcription studies in Corynebacterium glutamicum, we have developed an in vitro transcription system for this bacterium used as an industrial producer of amino acids and a model organism for actinobacteria. This system consists of C. glutamicum RNA polymerase (RNAP) core (alpha 2, beta, beta'), a sigma factor and a promoter-carrying DNA template, that is specifically recognized by the RNAP holoenzyme formed. The RNAP core was purified from the C. glutamicum strain with the modified rpoCgene, which produced His-tagged beta' subunit. The C. glutamicum sigA and sigH genes were cloned and overexpressed using the Escherichia coli plasmid vector, and the sigma subunits sigma(A) and sigma(H) were purified by affinity chromatography. Using the reconstituted C. glutamicum holo-RNAPs, recognition of the sigma(A)- and sigma(H)-dependent promoters and synthesis of the specific transcripts was demonstrated. The developed in vitro transcription system is a novel tool that can be us

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EE - Mikrobiologie, virologie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2012

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Applied Microbiology and Biotechnology

ISSN

0175-7598

e-ISSN

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Svazek periodika

96

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

9

Strana od-do

521-529

Kód UT WoS článku

000308953100022

EID výsledku v databázi Scopus

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