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Fig. 3 | Bioresources and Bioprocessing

Fig. 3

From: Zymomonas mobilis as an emerging biotechnological chassis for the production of industrially relevant compounds

Fig. 3

Genetic engineering tools to modify Zymomonas mobilis: A Plasmid-based approaches: Plasmids containing broad-host range replication origins can be maintained in Z. mobilis or shuttle vectors having Z. mobilis replication origin and an E. coli replication origin can be used instead; B Genome integration can be achieved in Z. mobilis via homologous recombination (HR) using a suicide vector, that is a plasmid lacking a suitable replication origin, or by using a plasmid expressing recombinases to catalyze HR and linear DNA fragments as donor template; C Genome editing mediated by Clustered Regularly Short Palindromic Repeats-associated Cas (CRISPR-Cas) systems: Heterologous CRISPR-Cas9 systems can be efficiently expressed in Z. mobilis to generate a double strand break (DSB) in genome followed by repair by HR by donor template. Here, a plasmid carrying a heterologous Cas9 protein and the guide RNA (gRNA) components is co-transformed with the homologous donor DNA fragment. Otherwise, the endogenous CRISPR-Cas system can be programmed to produce a specific DSB to be repaired by HR. Thereunto, a plasmid only carrying the gRNA elements is co-transformed with the homologous donor DNA fragment

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