Comprehensive Investigation of DNA Evolution in Enucleated Oocytes: Dual Functionality of PhiC31 and pCMV-LacZBbsI Vectors and Nonlinear Mutation Dynamics (A Simulation and Theoretical Study)
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2025
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| _version_ | 1866902084154032128 |
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| author | Reza Hashemi |
| author_facet | Reza Hashemi |
| contents | <p>Enucleated oocytes, lacking nuclear DNA and repair mechanisms, provide a unique system to explore<br>cytoplasmic influences on DNA evolution. This theoretical study investigates the transformation of two<br>separate plasmids—PhiC31 (3895 bp) and pCMV-LacZ-BbsI (4968 bp)—injected simultaneously into a<br>nucleus-free environment, with a combined total of 8863 base pairs. The vectors serve dual roles: confirming<br>nuclear absence through the silence of nuclear-dependent markers (KanR/NeoR, GFP, LacZ) and quantifying<br>mutation rates using a nonlinear model tailored for double-stranded DNA (dsDNA). Simulations predict a<br>baseline mutation rate of 4.47×10⁻⁶ mutations per nucleotide (0.040 mutations across 8863 bp) over 48 hours,<br>escalating to 1.00×10⁻⁴ (0.89 mutations) under high oxidative stress and ion interactions across 20 batches (5<br>cycles each, 240 hours per batch). Elevated temperature (T=40°C) and pressure (P=10 atm) amplify these<br>effects. We propose an experimental framework to validate these findings by injecting both plasmids<br>simultaneously into 100 enucleated mouse oocytes under varying conditions, offering insights into<br>cytoplasmic-driven DNA evolution with potential evolutionary implications.</p> |
| format | Recurso digital |
| id | zenodo_https___doi_org_10_5281_zenodo_15042327 |
| institution | Zenodo |
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| publishDate | 2025 |
| publisher | Zenodo |
| record_format | zenodo |
| spellingShingle | Comprehensive Investigation of DNA Evolution in Enucleated Oocytes: Dual Functionality of PhiC31 and pCMV-LacZBbsI Vectors and Nonlinear Mutation Dynamics (A Simulation and Theoretical Study) Reza Hashemi <p>Enucleated oocytes, lacking nuclear DNA and repair mechanisms, provide a unique system to explore<br>cytoplasmic influences on DNA evolution. This theoretical study investigates the transformation of two<br>separate plasmids—PhiC31 (3895 bp) and pCMV-LacZ-BbsI (4968 bp)—injected simultaneously into a<br>nucleus-free environment, with a combined total of 8863 base pairs. The vectors serve dual roles: confirming<br>nuclear absence through the silence of nuclear-dependent markers (KanR/NeoR, GFP, LacZ) and quantifying<br>mutation rates using a nonlinear model tailored for double-stranded DNA (dsDNA). Simulations predict a<br>baseline mutation rate of 4.47×10⁻⁶ mutations per nucleotide (0.040 mutations across 8863 bp) over 48 hours,<br>escalating to 1.00×10⁻⁴ (0.89 mutations) under high oxidative stress and ion interactions across 20 batches (5<br>cycles each, 240 hours per batch). Elevated temperature (T=40°C) and pressure (P=10 atm) amplify these<br>effects. We propose an experimental framework to validate these findings by injecting both plasmids<br>simultaneously into 100 enucleated mouse oocytes under varying conditions, offering insights into<br>cytoplasmic-driven DNA evolution with potential evolutionary implications.</p> |
| title | Comprehensive Investigation of DNA Evolution in Enucleated Oocytes: Dual Functionality of PhiC31 and pCMV-LacZBbsI Vectors and Nonlinear Mutation Dynamics (A Simulation and Theoretical Study) |
| url | https://doi.org/10.5281/zenodo.15042327 |