uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Building an Inducible T7 RNA Polymerase/T7 Promoter Circuit in Synechocystis sp. PCC6803
Carnegie Inst Sci, Dept Plant Biol, Stanford, CA 94305 USA.ORCID iD: 0000-0001-7460-4374
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Molecular Biomimetics.ORCID iD: 0000-0001-7256-0275
Carnegie Inst Sci, Dept Plant Biol, Stanford, CA 94305 USA.
2019 (English)In: ACS Synthetic Biology, E-ISSN 2161-5063, Vol. 8, no 4, p. 655-660Article in journal (Refereed) Published
Abstract [en]

To develop tightly regulated orthogonal gene expression circuits in the photoautotrophic cyanobacterium Synechocystis sp. PCC6803 (Syn6803), we designed a circuit in which a native inducible promoter drives the expression of phage T7 RNA polymerase (T7RNAP). T7RNAP, in turn, specifically recognizes the T7 promoter that is designed to drive GFP expression. In Syn6803, this T7RNAP/T7promoter-GFP circuit produces high, GFP fluorescence, which was further enhanced by using mutant T7 promoters. We also tested two orthogonal inducible promoters, Trc10 and L03, but these promoters drive T7RNAP to levels that are toxic in E. coli. Introduction of a protein degradation tag alleviated this problem. However, in Syn6803, these circuits did not function successfully. This highlights the underappreciated fact that similar circuits work with varying efficiencies in different chassis organisms. This lays the groundwork for developing new orthogonally controlled phage RNA polymerase-dependent expression systems in Syn6803.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC , 2019. Vol. 8, no 4, p. 655-660
Keywords [en]
Synechocystis sp. PCC6803, T7 RNA polymerase/T7 promoter circuit, orthogonal promoter, nickel inducible promoter
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-383202DOI: 10.1021/acssynbio.8b00515ISI: 000465487100006PubMedID: 30935196OAI: oai:DiVA.org:uu-383202DiVA, id: diva2:1338537
Available from: 2019-07-23 Created: 2019-07-23 Last updated: 2019-07-23Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMed

Authority records BETA

Lindblad, Peter

Search in DiVA

By author/editor
Jin, HaojieLindblad, Peter
By organisation
Molecular Biomimetics
In the same journal
ACS Synthetic Biology
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 15 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf