Evaluation of an E. coli Cell Extract Prepared by Lysozyme-Assisted Sonication and Transcriptional Interference in Toehold Switch-Based Circuits
Translated title:
Evaluation eines mit Hilfe von Lysozym- und Ultraschallbehandlung hergestellten E. coli-Zellextraktes und Transkriptionsinterferenz in Toehold-Switch-basierten Netzwerken
Author:
Falgenhauer, Elisabeth
Year:
2022
Document type:
Dissertation
Faculty/School:
TUM School of Natural Sciences
Advisor:
Simmel, Friedrich C. (Prof. Dr.)
Referee:
Simmel, Friedrich C. (Prof. Dr.); Gerland, Ulrich (Prof. Dr.)
Language:
en
Subject group:
PHY Physik
Keywords:
antisense RNA, transcriptional interference, gene regulation, bacterial extract, cell-free protein expression, phage assembly, mass spectroscopy, proteomics
Genetic circuits can be studied either in vivo or within cell-free systems. One of the crucial steps during the preparation of cell extract is the cell lysis procedure. In this study we prepared E. coli cell extract using a combination of lysozyme incubation and sonication cycles for the lysis step.
In a second project regulatory RNAs (toehold switches, triggers and anti-triggers) were studied within different promoter arrangements in vivo and transcriptional interference effects were analyzed.
«
Genetic circuits can be studied either in vivo or within cell-free systems. One of the crucial steps during the preparation of cell extract is the cell lysis procedure. In this study we prepared E. coli cell extract using a combination of lysozyme incubation and sonication cycles for the lysis step.
In a second project regulatory RNAs (toehold switches, triggers and anti-triggers) were studied within different promoter arrangements in vivo and transcriptional interference effects were analyzed...
»
Translated abstract:
Genetische Netzwerke können in vivo oder in zellfreien Systemen untersucht werden. Ein kritischer Schritt bei der Herstellung von Zellextrakt ist die Zelllyse. In dieser Arbeit wurde E. coli-Zellextrakt mit Hilfe einer Kombination aus Lysozyminkubation und Ultraschallzyklen hergestellt.
In einem zweiten Projekt wurden regulatorische RNAs (Toehold-Switches, Trigger und Anti-Trigger) innerhalb von verschiedenen Promotoranordnungen untersucht, wobei Transkriptionsinterferenz auftritt.