Cristin-resultat-ID: 1618160
Sist endret: 12. august 2019, 20:12

A new versatile shuttle vector for genome engineering of Bacillus subtilis by the CRISPR/Cas9 system

  • Antonio Garcia-Moyano og
  • Gro Elin Kjæreng Bjerga


Navn på arrangementet: Genome Engineering and Synthetic Biology
Sted: Bruges
Dato fra: 25. januar 2018
Dato til: 26. januar 2018


Arrangørnavn: Vlaams Instituut voor Biotechnologie

Om resultatet

Publiseringsår: 2018





Genredigering • Rekombinasjon, genetisk / genetisk rekombinasjon • Syntetisk biologi

Beskrivelse Beskrivelse


A new versatile shuttle vector for genome engineering of Bacillus subtilis by the CRISPR/Cas9 system


The adaptation of CRISPR/Cas9 technology to bacterial genetics facilitates manipulation of bacterial genomes ad libitum. The well-known gram-positive bacterium Bacillus subtilis is an attractive host for the production of heterologous proteins and it has therefore been object of some of the recent developments in bacterial genome engineering by CRISPR/Cas9 technology (Zhang et al., 2016; Altenbuchner, 2017). The different editing systems recently developed are based on a single broad host-range plasmid for creating mutant strains with improved fermentation capabilities. Due to the need of specific target sequences and repair templates for homologous recombination, obtaining the final knockout plasmid often requires several cloning steps. By means of restriction-free cloning techniques, we have adapted an existing shuttle plasmid by introducing a counter-selection cassette compatible with fragment exchange (FX) cloning technology (Geertsma and Dutzler, 2011). This versatile vector allows rapid and effective generation of a knockout plasmid for transformation and genome engineering in B. subtilis. In order to test this technology we have targeted the spoIIIAC gene. The gene encodes a sigma factor which has an essential role in controlling sporulation, allowing us to have an efficient read-out, as the lack of spores can be detected by xxx. We disrupted the gene in two related B. subtilis strains, rendering them asporogeneous. This new powerful tool provides some important advantages for targeted genome editing in an industrially relevant bacterial strain.


Antonio Garcia-Moyano

  • Tilknyttet:
    ved NORCE Norwegian Research Centre AS
Aktiv cristin-person

Gro Elin Kjæreng Bjerga

  • Tilknyttet:
    ved NORCE Klima og miljø ved NORCE Norwegian Research Centre AS
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