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

• 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
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.20578

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.