Efficient replication and generation of recombinant bovine adenovirus‐3 in nonbovine cotton rat lung cells expressing I‐SceI endonuclease
Bibliographic record
Abstract
BACKGROUND: The rigorous evaluation of recombinant bovine adenovirus (BAdV)-3 as a gene delivery vector requires quick and efficient method of isolating recombinants. This requires both a suitable cell line and an efficient method of rescuing recombinant BAdV-3. To facilitate rapid isolation of recombinant BAdV-3, we have developed an efficient system for generating recombinants using newly identified nonbovine cell line permissive for replication of BAdV-3. METHODS: Nonbovine cotton rat lung (CRL) cells in comparison to Madin-Darby bovine kidney cells and VIDO R2 cells were analyzed for the production of progeny virus and DNA transfection efficiency. In addition, lentiviral expression system was used to generate stable nonbovine CRL cell line expressing endonuclease I-SceI as examined by western blotting. Transfection of this cell line with circular or linear plasmid containing full-length BAdV-3 genome was used to generate recombinant BAdV-3. RESULTS: We demonstrate that nonbovine CRL cells are permissive for replication of BAdV-3 and can be efficiently transfected with plasmid DNA. Second, we constructed CRL cell line (VIDO DT1) expressing an intron-encoding endonuclease I-SceI. Finally, we demonstrate that transfection of VIDO DT1 cells with a circular plasmid containing recombinant BAdV-3 genome flanked by I-SceI recognition sites can efficiently rescue recombinant virus. CONCLUSIONS: The use of circular molecular clones together with I-SceI endonuclease expressing, BAdV-3 permissive CRL cell line not only increased the viral genome transfection efficiency, but also reduced the viral rescue time and amount of DNA required for rescuing recombinant BAdV-3s.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".