Nuclear transformation of a dinoflagellate symbiont of corals
Bibliographic record
Abstract
Dinoflagellates are a diverse and ecologically important group of single-celled eukaryotes. Many are photosynthetic autotrophs while others are predatory, parasitic, or symbiotic. One major group — the Symbiodiniaceae — is well known for its role as coral symbionts that provide the coral host with vital nutrients. While genetic transformation protocols have been published for some non-symbiotic dinoflagellate species, robust methods for genetic manipulation of coral symbionts are lacking, hindering a detailed molecular understanding of this critical symbiotic interaction. Here, we describe the successful transformation of coral symbiont Breviolum minutum (strain SSB01). Using Golden Gate modular plasmid assembly and electroporation, we drove transient NLS-GFP expression from an endogenous dinoflagellate virus nuclear protein (DVNP) promoter and successfully targeted GFP to the dinoflagellate nucleus. We further determined that puromycin can efficiently select transformed cells using the puromycin N-acetyltransferase ( pac ) resistance gene. Transformed cells could be maintained under antibiotic selection for at least 12 months without losing resistance, albeit with slowly attenuating fluorescence signal. We thus tested the expression of hybrid GFP-2A-PAC polypeptides under the control of a single promoter sequence to overcome loss of fluorescence, but lack of efficient 2A cleavage seemingly hindered antibiotic selection interfering GFP function. Despite this, our transformation approach now allows unanswered questions of dinoflagellate biology to be addressed, as well as fundamental aspects of dinoflagellate-coral symbiosis.
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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 | 0.000 | 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 | 0.001 |
| 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".