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1 Ιαν 2023 · The aim of this chapter is to provide an in-depth series of methods that will enable researchers to perform genetic manipulation of several different strains of B. subtilis. We describe five methods for genetic engineering: 1. Traditional gene manipulation, 2.
- Genome Editing Methods for Bacillus subtilis - PubMed
Use of the methods outlined in this chapter will allow...
- Genome Editing Methods for Bacillus subtilis - PubMed
Use of the methods outlined in this chapter will allow researchers to create gene insertions, deletions, substitutions, and RNA interference strains through a variety of methods custom to each application. Keywords: Bacillus subtilis; Genetic manipulation; Gram-positive; CRISPR/Cas9; Genetic engineering. © 2022.
19 Μαΐ 2022 · Bacillus subtilis is a widely studied Gram-positive bacterium that serves as an important model for understanding processes critical for several areas of biology including biotechnology and human health.
15 Αυγ 2016 · Here, we present a single-plasmid system which allows efficient genome editing of Bacillus subtilis. The plasmid pJOE8999 is a shuttle vector that has a pUC minimal origin of replication for Escherichia coli, the temperature-sensitive replication origin of plasmid pE194 ts for B. subtilis, and a kanamycin resistance gene working in both organisms.
29 Σεπ 2018 · We reviewe the design strategies for the development of efficient guide-RNAs and Cas9 in the CRISPR system for genome editing in B. subtilis. Finally, this review provides new perspectives on the use of CRISPR/Cas9 based genome engineering for sequence-mediated genome editing in B. subtilis.
31 Οκτ 2021 · The toolbox includes tools, such as double-strand and single-strand cutting CRISPR for point mutation, gene insertion, and gene deletion up to 38 kb. Moreover, catalytic dead Cas proteins have been used for base editing, as well as for the control of gene expression (CRISPRi and CRISPRa).
6 Μαΐ 2019 · This method shortens the overall process from 1 week to 1 day and allows the integration of multiple genes in one step, providing a simple and fast method for genome editing in B. subtilis. Keywords: DNA assembly; gene knockout/knockin; genome editing mutant libraries; transformation efficiency.
