ABSTRACT
Bacteriophages-viruses that infect bacterial cells - are the most abundant biological entities on Earth. The use of phages in fundamental research and industry requires tools for precise manipulation of their genomes. Yet, compared to bacterial genome engineering, modification of phage genomes is challenging because of the lack of selective markers and thus requires laborious screenings of recombinant/mutated phage variants. The development of the CRISPR-Cas technologies allowed to solve this issue by the implementation of negative selection that eliminates the parental phage genomes. In this manuscript, we summarize current methods of phage genome engineering and their coupling with CRISPR-Cas technologies. We also provide examples of our successful application of these methods for introduction of specific insertions, deletions, and point mutations in the genomes of model Escherichia coli lytic phages T7, T5, and T3.
Subject(s)
Bacteriophages , Bacteriophage T7/geneticsABSTRACT
Stereotypy and locomotor hyperactivity caused by subcutaneous injection of apomorphine (1-5 mg) were quantitated with an "Animex-DSE" motility meter. Intraperitoneal administration of galantamin in a dose of 2.5 mg/kg 10 minutes before apomorphine reversed the latter's effect. However, when injected in a similar way 40 minutes before, it exerted no action on behavioral effects of apomorphine. Intraperitoneal administration of amizyl in a dose of 1 mg/kg completely prevented antagonism of galantamin to apomorphine activity but did not potentiate its effect on the locomotor activity in the animals. The data obtained show that the effect of the cholinergic system on dopaminergic mediation in individual brain structures may be modulating in nature. The experimental data are discussed in the light of the present-day concepts of the function of dopamine receptors in the striatum and mesolimbic systems of the brain.
