After all the components listed are released into the packaging cells, viral proteins and recombinant RN A guaranteeing the formation Vortioxetine of the HIV 1 like particles that are released into the environment are synthesized inside the afore-mentioned cells. The inclusion of those particles to the target cells induces the synthesis of the DNA of a provirus that contains a marker gene, whose integration in to the target cell genome renders it able to fluorescing around the recombinant RN A genome in target cells. It must be stressed that the usage of plasmid DNAs expressing individual virus specific proteins enables to make any versions of pseudo HIV 1 particles with one or several variations in any chemical of viral replication which correspond to the drug-resistant HIV 1 strains. So far, revealed investigations still contain an insufficient amount of cases locomotor system of effective use of these systems to review the antiretroviral activity of substances that differ in their character, this causes it to be unclear exactly how common the described systems are. In this regard, our study primarily endeavoured to verify the adequacy of the cell system recommended for testing potential anti-hiv 1 agencies. The game of several of inhibitors of HIV 1 reverse transcriptase and integrase were examined, both of which have found application in medical practice and have withstood different phases of laboratory research. EXPERIMENTAL Cell expansion These cell lines were used in this study: HEK293, SC 1, Jurkat, CE Michael SS, and Kasumi 1.. The HEK293 and SC 1 cell lines were cultured in DMEM containing Everolimus ic50 one hundred thousand fetal calf serum, 4 mM of L glutamine, 100 U /ml of penicillin, and 100 ug/ml of streptomycin. . The CE M Empire Simba, Jurkat, and Kasumi 1 cell lines were cultured in RPMI 1640 containing mM of L glutamine, 4 2000-2009 FCS, 100 U /ml of penicillin, and 100 ug/ml of streptomycin.. The cells were grown at 37 in moist air containing five hundred of 2.. Obtainment of pseudo HIV 1 particles HEK293 cells seeded in Petri dishes with a diameter of 100 mm inside the number of 3. 0 3. 5 106 cells per dish 12 14 h prior to the transfection on-set were used as packaging cells, when the construction of recombinant lentiviral particles occurs. DNA of the lentiviral vector containing the marker gene of green fluorescent protein and the plasmids directing the synthesis of the proteins which are necessary for the formation of pseudo HIV 1 particles were introduced into HEK293 cells via calcium phosphate transfection. The infectious pseudo HIV 1 particles were obtained 24 h following transfection using a 12 h interval. The virus was titrated on HEK293 cells seeded to 24 well plates 24 h before infection. The degree of mobile fluorescence was measured on an Epics 4XL Beckman Coulter move cytofluorimeter 48 h after the infection.
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