Publication detail

Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum

KOLEK, J. SEDLÁŘ, K. PROVAZNÍK, I. PATÁKOVÁ, P.

Original Title

Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum

English Title

Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum

Type

conference proceedings

Language

en

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.

English 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

Released

25.04.2016

Location

Mikulov

ISBN

978-80-86238-91-3

Book

4. mezinárodní chemicko-technologická konference SBORNÍK ABSTRAKT A PLNÝCH TEXTŮ

Pages from

1

Pages to

1

Pages count

1

BibTex


@proceedings{BUT124791,
  author="Jan {Kolek} and Karel {Sedlář} and Ivo {Provazník} and Petra {Patáková}",
  title="Development of Methods for Genetic Manipulation of Solventogenic, Non-Type Strain Clostridium Pasteurianum",
  annote="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.
",
  booktitle="4. mezinárodní chemicko-technologická konference SBORNÍK ABSTRAKT A PLNÝCH TEXTŮ",
  chapter="124791",
  howpublished="electronic, physical medium",
  year="2016",
  month="april",
  pages="1--1",
  type="conference proceedings"
}