Mg<sup>2+</sup>-free buffer elevates transformation efficiency of<i>Vibrio parahaemolyticus</i>by electroporation
Bibliographic record
Abstract
AIMS: The ability to transform Vibrio spp. is limited by the extracellular nuclease that their cells secrete. The reported transformation efficiency of this organism is 10(2)-10(5) transformants per microgram DNA. We tried different buffers and conditions, aiming to elevate its transformation efficiency. METHODS AND RESULTS: MgCl(2) and sucrose are often included in the washing and/or electroporation buffers to stabilize the cell membrane. However, Mg(2+) is required for production and activity of the extracellular nuclease. A simple electroporation buffer lacking Mg(2+) was found to increase transformation efficiency dramatically, to levels 50-fold more than the buffers containing Mg(2+). To maintain the stability of the cell membranes, Mg(2+) was replaced with high concentrations of sucrose, from 272 to 408 mmol l(-1). With the new buffers, the transformation efficiency of Vibrio parahaemolyticus was increased to 2.2 x 10(6) transformants per microgram DNA. CONCLUSIONS: Mg(2+) in the buffer adversely affected transformation of V. parahaemolyticus by electroporation. The cell membranes of vibrio can be stabilized by high concentration of sucrose when Mg(2+) is absent. SIGNIFICANCE AND IMPACT OF THE STUDY: A greater transformation efficiency can facilitate the genetic analysis of an organism and its pathogenicity. Buffers lacking Mg(2+) can be used for other nuclease-producing organisms.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| 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 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
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 teacher head, 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".