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To investigate the role of each gene of pilus island 2a (Figure 6A) in biofilm formation, we also analyzed biofilm formation in the mutant strains, generated in strain 515, lacking either one of the three structural LPXTG proteins (BP, AP1 and AP2) or the two SrtC-like sortases SrtC-1 and SrtC-2 [22].
Intriguingly in the Δ Bar1 mutant strains generated in this study, sporulation resulted in the formation of shorter, less uniform spores with fewer cells than the WT (Rittenour et al., 2011).
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The roles of the various mechanisms were also investigated by simulating the stochastic model on in silico mutant strains generated by eliminating individual mechanisms.
We employed both C. graminicola wild type (WT) strain CgM2 and mutant strains generated by Agrobacterium tumefaciens-mediated transformation (ATMT), which are affected in virulence to different extent.
Six independent ΔEZL3 mutant strains were generated in each of the B2086 and CU428 backgrounds (that are compatible for mating).
Three of the mutant strains we generated in this study, CN579, CN1162 and CN1574 carried two point mutations within the target gene.
Mutant strains were generated with a dose of 4-NQO resulting in 97% kill.
Mutant strains were generated by transposon mutagenesis.
All mutant strains were generated by gene SOEing [ 34].
In this study, an unmarked ΔpurMCD mutant was generated in type A strain Schu S4 and its ability to confer protective immunity against wild-type F. tularensis was investigated.
As an alternative, a mutant was generated in the G27 strain whose ansB gene (HPG27_679) had been previously isolated and which shows close homology with the CCUG 17874 ansB gene (HP0723) [30] (Supplementary Fig. S2).
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