---
_id: '11713'
abstract:
- lang: eng
  text: "Objective: MazF is a sequence-specific endoribonuclease-toxin of the MazEF
    toxin–antitoxin system. MazF cleaves single-stranded ribonucleic acid (RNA) regions
    at adenine–cytosine–adenine (ACA) sequences in the bacterium Escherichia coli.
    The MazEF system has been used in various biotechnology and synthetic biology
    applications. In this study, we infer how ectopic mazF overexpression affects
    production of heterologous proteins. To this end, we quantified the levels of
    fluorescent proteins expressed in E. coli from reporters translated from the ACA-containing
    or ACA-less messenger RNAs (mRNAs). Additionally, we addressed the impact of the
    5′-untranslated region of these reporter mRNAs under the same conditions by comparing
    expression from mRNAs that comprise (canonical mRNA) or lack this region (leaderless
    mRNA).\r\nResults: Flow cytometry analysis indicates that during mazF overexpression,
    fluorescent proteins are translated from the canonical as well as leaderless mRNAs.
    Our analysis further indicates that longer mazF overexpression generally increases
    the concentration of fluorescent proteins translated from ACA-less mRNAs, however
    it also substantially increases bacterial population heterogeneity. Finally, our
    results suggest that the strength and duration of mazF overexpression should be
    optimized for each experimental setup, to maximize the heterologous protein production
    and minimize the amount of phenotypic heterogeneity in bacterial populations,
    which is unfavorable in biotechnological processes."
acknowledgement: "We acknowledge the Max Perutz Labs FACS Facility together with Thomas
  Sauer. NN is grateful to Călin C. Guet for his support.\r\nThis work was funded
  by the Elise Richter grant V738 of the Austrian Science Fund (FWF), and the FWF
  Lise Meitner grant M1697, to NN; and by the FWF grant P22249, FWF Special Research
  Program RNA-REG F43 (subproject F4316), and FWF doctoral program RNA Biology (W1207),
  to IM. Open access funding provided by the Austrian Science Fund."
article_number: '173'
article_processing_charge: No
article_type: letter_note
author:
- first_name: Nela
  full_name: Nikolic, Nela
  id: 42D9CABC-F248-11E8-B48F-1D18A9856A87
  last_name: Nikolic
  orcid: 0000-0001-9068-6090
- first_name: Martina
  full_name: Sauert, Martina
  last_name: Sauert
- first_name: Tanino G.
  full_name: Albanese, Tanino G.
  last_name: Albanese
- first_name: Isabella
  full_name: Moll, Isabella
  last_name: Moll
citation:
  ama: Nikolic N, Sauert M, Albanese TG, Moll I. Quantifying heterologous gene expression
    during ectopic MazF production in Escherichia coli. <i>BMC Research Notes</i>.
    2022;15. doi:<a href="https://doi.org/10.1186/s13104-022-06061-9">10.1186/s13104-022-06061-9</a>
  apa: Nikolic, N., Sauert, M., Albanese, T. G., &#38; Moll, I. (2022). Quantifying
    heterologous gene expression during ectopic MazF production in Escherichia coli.
    <i>BMC Research Notes</i>. Springer Nature. <a href="https://doi.org/10.1186/s13104-022-06061-9">https://doi.org/10.1186/s13104-022-06061-9</a>
  chicago: Nikolic, Nela, Martina Sauert, Tanino G. Albanese, and Isabella Moll. “Quantifying
    Heterologous Gene Expression during Ectopic MazF Production in Escherichia Coli.”
    <i>BMC Research Notes</i>. Springer Nature, 2022. <a href="https://doi.org/10.1186/s13104-022-06061-9">https://doi.org/10.1186/s13104-022-06061-9</a>.
  ieee: N. Nikolic, M. Sauert, T. G. Albanese, and I. Moll, “Quantifying heterologous
    gene expression during ectopic MazF production in Escherichia coli,” <i>BMC Research
    Notes</i>, vol. 15. Springer Nature, 2022.
  ista: Nikolic N, Sauert M, Albanese TG, Moll I. 2022. Quantifying heterologous gene
    expression during ectopic MazF production in Escherichia coli. BMC Research Notes.
    15, 173.
  mla: Nikolic, Nela, et al. “Quantifying Heterologous Gene Expression during Ectopic
    MazF Production in Escherichia Coli.” <i>BMC Research Notes</i>, vol. 15, 173,
    Springer Nature, 2022, doi:<a href="https://doi.org/10.1186/s13104-022-06061-9">10.1186/s13104-022-06061-9</a>.
  short: N. Nikolic, M. Sauert, T.G. Albanese, I. Moll, BMC Research Notes 15 (2022).
date_created: 2022-08-01T09:04:27Z
date_published: 2022-05-13T00:00:00Z
date_updated: 2022-08-01T09:27:40Z
day: '13'
ddc:
- '570'
department:
- _id: CaGu
doi: 10.1186/s13104-022-06061-9
external_id:
  pmid:
  - '35562780'
file:
- access_level: open_access
  checksum: 008156e5340e9789f0f6d82bde4d347a
  content_type: application/pdf
  creator: dernst
  date_created: 2022-08-01T09:24:42Z
  date_updated: 2022-08-01T09:24:42Z
  file_id: '11714'
  file_name: 2022_BMCResearchNotes_Nikolic.pdf
  file_size: 1545310
  relation: main_file
  success: 1
file_date_updated: 2022-08-01T09:24:42Z
has_accepted_license: '1'
intvolume: '        15'
keyword:
- General Biochemistry
- Genetics and Molecular Biology
- General Medicine
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 26956E74-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: V00738
  name: Bacterial toxin-antitoxin systems as antiphage defense mechanisms
publication: BMC Research Notes
publication_identifier:
  issn:
  - 1756-0500
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
related_material:
  link:
  - relation: erratum
    url: https://doi.org/10.1186/s13104-022-06152-7
scopus_import: '1'
status: public
title: Quantifying heterologous gene expression during ectopic MazF production in
  Escherichia coli
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 15
year: '2022'
...