Cristin-resultat-ID: 1618160
Sist endret: 12. august 2019, 20:12
Resultat
Poster
2018

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

Bidragsytere:
  • Antonio Garcia-Moyano og
  • Gro Elin Kjæreng Bjerga

Presentasjon

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

Arrangør:

Arrangørnavn: Vlaams Instituut voor Biotechnologie

Om resultatet

Poster
Publiseringsår: 2018

Klassifisering

Vitenskapsdisipliner

Molekylærbiologi

Emneord

Genredigering • Rekombinasjon, genetisk / genetisk rekombinasjon • Syntetisk biologi

Beskrivelse Beskrivelse

Tittel

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

Sammendrag

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.

Bidragsytere

Antonio Garcia-Moyano

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

Gro Elin Kjæreng Bjerga

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