Fig. 3: Results of single and multiple deletions with NT-CRISPR.
a Table providing information about deleted sequence. Locus tags of deleted genes are as follows: Pili (PN96_01310 - PN96_01315), flagella (PN96_02540 - PN96_02685), vnp1 (PN96_04290 - PN96_04520), vnp2 (PN96_06880 - PN96_07085), galE (PN96_22140), xds(PN96_19285). Chr1 = Chromosome 1, Chr2 = Chromosome 2. b Efficiency of deletions. Positive colonies were identified by PCR assays (n = 50 colonies). The dashed line indicates the highest possible value. c Visualization of NT-CRISPR plasmid carrying three gRNAs. Colored squares indicate matching fusion sites used for construction of this plasmid using Golden Gate Assembly. More details regarding the assembly of a multi gRNA NT-CRISPR plasmid is provided in Supplementary Fig. S5a,b. SBOL symbols for omitted detail (three points) represent the transcriptional unit for the regulatory proteins LacI and TetR for Ptac and Ptet, respectively. d, e Venn diagrams to visualize results of multigene deletions. Note that areas of ellipses and intersections are not proportional to the displayed values. Colonies of cells carrying none of the deletions are indicated with a separate ellipsis (gray). Results were obtained by PCR assays (n = 50 colonies). Plasmids used are pST_138 (d) and pST_137 (e).
