Jürgen Britta, Barken Kim Bundvig, Tobisch Steffen, Pioch Daniel, Wümpelmann Mogens, Hecker Michael, Schweder Thomas
Institute of Microbiology, Ernst-Moritz-Arndt-University Greifswald, F.-L.-Jahnstrasse 15, D-17487 Greifswald, Germany.
Biotechnol Bioeng. 2005 Nov 5;92(3):299-307. 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.
关键过程相关基因的知识可用于改进生物过程的控制。到目前为止,与生物过程相关的标记基因只能通过既定的表达分析方法进行离线分析。在本研究中,提出了一种在生物过程中对基因表达进行潜在在线监测的替代方法。该方法基于在电DNA芯片上结合基于磁珠的夹心杂交对特定mRNA进行测量。为了实现对特定mRNA的在线测量,开发了一种快速且部分自动化分离高质量RNA样本的改进方法。将电DNA芯片的表达分析与光学DNA微阵列以及实时RT-PCR对枯草芽孢杆菌三个选定的过程相关基因进行了比较。我们证明,通过电DNA芯片进行的mRNA分析与微阵列分析和实时RT-PCR技术相比,结果相似。