Cloning and expression of a neutral protein gene BS2 from Bacillus subtilis B111 in Pichia pastoris.
Bibliographic record
Abstract
【Objective】 In order to exploit novel disease-resistance gene and biological pesticide sources,antifungal protein secreted by Bacillus subtilis B111 was purified and identified,then its gene was cloned and expressed in Pichia pastoris.【Method】 Antifungal protein was purified by ultrafiltration,90% ammonium sulfate precipitation and QAE Sephadex A50 ion exchange chromatogram column,then it was identified via CapLC-ESI-MS-MS.Its gene encoding antifungal protein was cloned.The recombinant expression vector pPIC9K-PT was constructed and transformed into P.astoris GS115.The transformants were identified by PCR analysis.The products of the transformants induced by methanol were identified by neutral protease and antifungal activity assay.【Result】 An antifungal protein BS2 was purified and identified to be a neutral protease from B.subtilis.Antifungal gene BS2 with pro-sequence was cloned.The recombinant expression vector pPIC9K-PT was constructed and BS2 gene was expressed in P.pastoris.The recombinant BS2 protein exhibited a molecular mass of approximately 69 kD and presented neutral protease activity of 27 800 U·g-1 and antifungal activity.Its production reached up to as high as 29.77 mg·L-1.【Conclusion】 In this study,an antifungal neutral protein BS2 was purified and identified,then its gene was cloned and successfully expressed in P.pastoris.
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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".