Optimization of methods of site-specific mutagenesis for Pseudomonas bacteria genus using pK18mob plasmid vector
Keywords:
site-specific insertion mutagenesis, suicide integrative vector, geno-engineering construction, transconjugantsAbstract
In the course of this study, a method for site-directed insertional mutagenesis was developed using the pK18mob suicide integral vector for bacteria of the genus Pseudomonas. It is shown that the optimal ratio of donor and recipient cells is 50 : 50. At the same time, in bacteria of P. chlororaphis, as well as their subspecies, the optimal ratio is shifted towards the predominance of donor cells. The use of this technique allows to increase the yield of transconjugants more than 4 times, which significantly expands the possibilities for searching and selecting the variants of strains of interest to the researcher. It was also found that the collection strains of bacteria of the genus Pseudomonas have different growth rates. It was demonstrated that the gene psrA is present in the genomes of all strains studied.
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