Přístupnostní navigace
E-application
Search Search Close
Publication detail
KOLEK, J. SEDLÁŘ, K. PROVAZNÍK, I. PATÁKOVÁ, P.
Original Title
Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum
Type
conference proceedings
Language
English
Original Abstract
The existence of methods for genetic manipulation of industrial microorganisms is generally essential for improving their properties like productivity or resistance against their own products. These methods are also very important for better, quicker and more effective research that could lead to the acquisition of important information useful in industrial processes. First step of every genetic manipulation is generally introduction of foreign DNA molecule into the bacterial cell and the most commonly used method for introducing DNA molecule into bacterial cells is transformation when exogenous molecule of DNA is introduced directly through the cell membrane. We described step-by-step development of methods for genetic modification of C. pasteurianum NRRL B-598 – a solventogenic bacterium producing butanol, acetone and ethanol which is unique in its exceptional oxygen resistance. Firstly, potential restriction modification systems, which could prevent transformation, were investigated. In the following steps, different transformation approaches and conditions were tested. It was obtained that unmethylated vector is necessary to use for good transformation efficiency and electroporation, conjugation with EscherichiaEshcerichia coli donor and sonoporation were used for transformation successfully. Sonoporation, method based on the ultrasound-mediated cavitation of bacterial membrane, was revealed as new, very efficient transformation technique for C. pasteurianum NRRL B-598. This method does not require any special equipment and is very fast, which could be very important parameter for anaerobic bacteria transformation.
Keywords
Clostridia; biotechnology
Authors
KOLEK, J.; SEDLÁŘ, K.; PROVAZNÍK, I.; PATÁKOVÁ, P.
Released
25. 4. 2016
Location
Mikulov
ISBN
978-80-86238-91-3
Book
4. mezinárodní chemicko-technologická konference SBORNÍK ABSTRAKT A PLNÝCH TEXTŮ
Pages from
1
Pages to
Pages count
BibTex
@proceedings{BUT124791, editor="Jan {Kolek} and Karel {Sedlář} and Valentine {Provazník} and Petra {Patáková}", title="Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum", year="2016", pages="1--1", address="Mikulov", isbn="978-80-86238-91-3" }