ABSTRACT
None of the studied eukaryotic species has a natural system for homologous recombination of the mitochondrial genome. We propose an integrated genetic construct pQ-SRUS, which allows introduction of the recA gene from Bacillus subtilis into the nuclear genome of an extremophilic yeast, Yarrowia lipolytica. The targeting of recombinant RecA to the yeast mitochondria is provided by leader sequences (5'-UTR and 3'-UTR) derived from the SOD2 gene mRNA, which exhibits affinity to the outer mitochondrial membrane and thus provides cotranslational transport of RecA to the inner space of the mitochondria. The Y. lipolytica strain bearing the pQ-SRUS construct has the unique ability to integrate DNA constructs into the mitochondrial genome. This fact was confirmed using a tester construct, pQ-NIHN, intended for the introduction of the EYFP gene into the translation initiation region of the Y. lipolytica ND1 mitochondrial gene. The Y. lipolytica strain bearing pQ-SRUS makes it possible to engineer recombinant producers based on Y. lipolytica bearing transgenes in the mitochondrial genome. They are promising for the construction of a genetic system for in vivo replication and modification of the human mitochondrial genome. These strains may be used as a tool for the treatment of human mitochondrial diseases (including genetically inherited ones).
Subject(s)
Bacterial Proteins/genetics , Genetic Engineering , Genome, Mitochondrial , Homologous Recombination , Rec A Recombinases/genetics , Yarrowia/genetics , Bacillus subtilis/genetics , Bacterial Proteins/biosynthesis , Humans , Rec A Recombinases/biosynthesis , Yarrowia/metabolismABSTRACT
The objective of the study is elucidation of perspectives of 3,3'-diathylcarbocyaine application as a photosensitizer for curing viral infections by photodynamic therapy. Lipid-containing bacteriophage PM-2 of Pseudoalteromonas espejiana was used as a model. The testing was carried out at a special installation modeling photodynamic exposure conditions towards a non-fractionated phage lysate. 3,3'-DECC demonstrated a rapid photo-bleaching when added tothe phage lysate but not to water. The initial rate of PM-2 phage photoinactivation was proportional to the square concentration of the dye in the range of 0.5-9 µmol/L. This confirms a hypothesis that the dimer is the principal photochemically active form of the dye. An improved ability to form dimers was found in the dye in the phage lysate (10-folds better than in the water). The dye formed a stable adduct with the bacteriophage material. This adduct had an extinction maximum at λ(max) = 594 nm and demonstrated the properties of a polymer (sedimentation under a low-speed centrifugation).
Subject(s)
Benzothiazoles/pharmacology , Carbocyanines/pharmacology , Corticoviridae/drug effects , Models, Biological , Photochemotherapy/methods , Photosensitizing Agents/pharmacology , Benzothiazoles/chemistry , Benzothiazoles/therapeutic use , Carbocyanines/chemistry , Carbocyanines/therapeutic use , Corticoviridae/radiation effects , Dimerization , Photosensitizing Agents/chemistry , Photosensitizing Agents/therapeutic use , Pseudoalteromonas/virology , Virus Diseases/drug therapy , Virus Diseases/radiotherapyABSTRACT
AIM: To study characteristics of oculomotor movements in schizophrenia. MATERIAL AND METHODS: Oculomotor characteristics of 30 patients with diagnosis of paranoid schizophrenia and 20 people of the control group were measured. RESULTS AND CONCLUSION: Parameters of specific eye movements, which are characteristic of this disease, have been identified. A comparison with the control group has been performed. It has been concluded that videooculography can be used as an additional method for examination of patients with schizophrenia.