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Rapid monitoring of mRNA levels with a molecular beacon during microbial fermentation

Paper ID Volume ID Publish Year Pages File Format Full-Text
24487 43518 2010 7 PDF Available
Title
Rapid monitoring of mRNA levels with a molecular beacon during microbial fermentation
Abstract

In the microbial fermentation bioreactor, the processes of mRNA transcription, protein translation, and enzyme-catalyzed biosynthesis remain as “black boxes” of industrial monitoring and process control. Monitoring the kinetics of these “black boxes” is very helpful for optimizing and controlling the microbial fermentation process. This study first applied a molecular beacon (MB) to monitor the changes in the mRNA level of the phzC gene during antibiotic phenazine-1-carboxylic acid fermentation. Seven typical MB hybridization buffers were compared, and the effect of formamide on MBs was also studied. The results showed that rapid monitoring of the mRNA level using MBs was feasible. The optimal hybridization buffer for phzC MB was 100 mM Tris, 1 mM MgCl2, pH 8.0. The optimal hybridization temperature was 35 °C, and formamide proved unsuitable for MB hybridization. The limit of detection of phzC MB was 1.67 nM and MB hybridization was complete by 7 min. Given that the time for RNA extraction is 12 min, it is possible that monitoring of phzC mRNA can be completed in less than 20 min. Since production of most amine acids, organic acids, wines, antibiotics, and proteins relies on microbial fermentation, our method may have some potential for application in these other microbial industries.

Keywords
Molecular beacon; mRNA monitoring; Hybridization condition; Phenazine-1-carboxylic acid
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Rapid monitoring of mRNA levels with a molecular beacon during microbial fermentation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 145, Issue 3, 1 February 2010, Pages 310–316
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us