Application of an electric DNA-chip for the expression analysis of bioprocess-relevant marker genes of Bacillus subtilis

The knowledge of critical process‐relevant genes can be used for an improved control of bioprocesses. So far bioprocess‐relevant marker genes can be analyzed by established expression analysis methods only off‐line. In this study, an alternative approach for a potential at‐line monitoring of gene ex...

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Published in:Biotechnology and bioengineering 2005-11, Vol.92 (3), p.299-307
Main Authors: Jürgen, Britta, Barken, Kim Bundvig, Tobisch, Steffen, Pioch, Daniel, Wümpelmann, Mogens, Hecker, Michael, Schweder, Thomas
Format: Article
Language:English
Subjects:
at-line monitoring
Bacillus subtilis
Bacillus subtilis - metabolism
Bacteria
Bacterial Proteins - metabolism
Biological and medical sciences
Biomarkers
bioprocess control
Biotechnology
Deoxyribonucleic acid
DNA
electric DNA-chip
Electrochemistry - methods
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression Profiling - methods
Genetic markers
In Situ Hybridization - methods
Methods. Procedures. Technologies
Microbial engineering. Fermentation and microbial culture technology
Oligonucleotide Array Sequence Analysis - methods
Online Systems
RNA isolation
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Summary:The knowledge of critical process‐relevant genes can be used for an improved control of bioprocesses. So far bioprocess‐relevant marker genes can be analyzed by established expression analysis methods only off‐line. In this study, an alternative approach for a potential at‐line monitoring of gene expression during bioprocesses is suggested. This approach is based on the measurement of specific mRNAs on an electric DNA‐chip in connection with a magnetic bead‐based sandwich hybridization. In order to allow an at‐line measurement of specific mRNAs an improved method for a fast and partially automated isolation of high quality‐RNA samples was developed. The expression analysis of the electric DNA‐chip was compared with optical DNA micro arrays and the real time RT‐PCR for three selected process‐relevant genes of Bacillus subtilis. We demonstrate that the mRNA analysis by means of the electric DNA‐chip gives similar results compared to the micro array analysis and the real time RT‐PCR technique. © 2005 Wiley Periodicals, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.20578