Formation of the D-loop structure complexes between DNA and oligonucleotides and affinity modyfication of DNA by chemically reactive derivatives of oligonucleotides lead to the recombinationOlga I Sinitsina, Konstantin V. Stoletov, Dmitry V. Bugreev, Galiya A. Maksakova, Irina F. Bugreeva, Elena A. Vasyunina, Georgy A. Nevinsky
Med Sci Monit 2000; 6(4): BR653-657 :: ID: 508090
Background: Affinity modification of DNA by chemically reactive derivatives of complementary oligonucleotides (ODNs) and antisense ODNs has shown an application for the inhibition of gene expression and the growth of viruses and parasites in high organisms. Unfortunately, the rapid advancement of antisense therapeutic approaches is not parallel to the investigation of possible consequences of antisense and gene-directed ODNs on genetic material of the cells being treated. Here we tried for the first time to estimate a possible genetic impact of antisense ODNs and their chemically reactive derivatives on the cells using bacteria and the plasmid DNA.
Material/Methods: Recombination of direct repeats, induced by the formation of reversible complexes of plasmid DNA with complementary ODNs and after covalent binding of the alkylating derivative of the ODNs with DNA, has been investigated. For this purpose, a polylinker sequence flanked by 165 bp direct repeats was inserted within the tet gene of pBR 327. This plasmid was used to construct DNA containing AT- and GC- rich sequences placed in the central region of the polylinker.
Results: Transformation of E. coli cells with the plasmids (and with mixtures of the plasmids with d(pN)17, complementary to the AT- and GC- rich sequences) did not produce deletions. After modification of plasmids with alkylating derivatives of d(pN)17,the deletion of the polylinker DNA region (recombination) revealed the restoration of the tet gene function. The same effect was found at the cell transformations with the D-loop complex of the plasmids with ODNs, but the frequency of the transformants was about 1.5-2 times lower. The data obtained demonstrate that the complexes of DNA with complementary ODNs and the modification of the plasmids by reactive ODN derivatives result in induction of the recombination process and in loss of genetic material.
Keywords: Bacteria, recombination, repair, plasmid DNA modification, alkylating oligonucleotide derivatives
01 May 2023 : EditorialEditorial: Twenty Years On from Sequencing the Human Genome, Personalized/Precision Oncology Prepares to Meet the Challenges of Checkpoint Inhibitor Therapy
Med Sci Monit 2023; 29:e940911
31 May 2023 : Review articlePrevalence, Diagnosis, and Management of Eclampsia and the Need for Improved Maternal Care: A Review
Med Sci Monit In Press; DOI: 10.12659/MSM.939919
30 May 2023 : Clinical ResearchA Case-Control Study of Cognitive Function in Patients with End-Stage Renal Disease Before and After Hemodi...
Med Sci Monit In Press; DOI: 10.12659/MSM.940409
30 May 2023 : Review articleA Review of the Role of the Zebrafish (Danio reiro) in Preclinical and Clinical Models of Biomarker Identif...
Med Sci Monit In Press; DOI: 10.12659/MSM.940550
30 May 2023 : Database AnalysisThe COVID-19 Crisis and the Incidence of Alcohol-Related Deaths in Poland
Med Sci Monit In Press; DOI: 10.12659/MSM.940904
Most Viewed Current Articles
13 Nov 2021 : Clinical ResearchAcceptance of COVID-19 Vaccination and Its Associated Factors Among Cancer Patients Attending the Oncology ...
Med Sci Monit 2021; 27:e932788
30 Dec 2021 : Clinical ResearchRetrospective Study of Outcomes and Hospitalization Rates of Patients in Italy with a Confirmed Diagnosis o...
Med Sci Monit 2021; 27:e935379
08 Mar 2022 : Review articleA Review of the Potential Roles of Antioxidant and Anti-Inflammatory Pharmacological Approaches for the Man...
Med Sci Monit 2022; 28:e936292
01 Jan 2022 : EditorialEditorial: Current Status of Oral Antiviral Drug Treatments for SARS-CoV-2 Infection in Non-Hospitalized Pa...
Med Sci Monit 2022; 28:e935952