A rapid CAT transformation protocol and nuclear transgene expression tools for metabolic engineering in Cyanidioschyzon merolae 10D
Bibliographic record
Abstract
The eukaryotic red alga Cyanidioschyzon merolae 10D is an emerging algal host for synthetic biology and metabolic engineering. Its small nuclear genome (16.5 Mb; 4775 genes), low intron content (38), stable transgene expression, and capacity for homologous recombination into its nuclear genome make it ideal for genetic and metabolic engineering endeavors. Here, we present an optimized transformation and selection protocol, which yields single chloramphenicol-resistant transformants in under two weeks. Transformation dynamics and a synthetic modular plasmid toolkit are reported, including several new fluorescent reporters. Techniques for fluorescence reporter imaging and analysis at different scales are presented to facilitate high-throughput screening of C. merolae transformants. We use this plasmid toolkit to overexpress the Ipomoea batatas isoprene synthase and demonstrate the dynamics of engineered volatile isoprene production during different light regimes using multi-port headspace analysis coupled to parallel photobioreactors. This work seeks to promote C. merolae as an algal system for metabolic engineering and future sustainable biotechnological production. • Protocol for nuclear genome transformation yields colonies in under 2 weeks • Multiple new fluorescent reporters and modular plasmid toolkit developed • Engineered isoprene produced from the alga • Isoprene quantified by parallel inline headspace analysis coupled to photobioreactors
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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) | 0.001 | 0.001 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.003 | 0.004 |
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".