Cloning Expression and Purification of Heat Shock Protein HSP60 from Hyperthermophilic Archaeon Pyrococcus furiosus
Bibliographic record
Abstract
[Objective] The aim was to study the cloning,expression and purification method of heat shock protein HSP60 from Hyperthermophilic Archaeon Pyrococcus furiosus.[Method] The homology analysis of Pfu-HSP60 gene sequence was taken by using Canada HSP60 database and sequence comparative tools of BLAST and FASTA.HSP60 gene was amplified in vitro and cloned and expressed in Escherichia coli.Pfu-HSP60 protein was also purified.[Result] With the Taq enzyme as DNA polymerase,the anticipative Pfu-HSP60 gene was amplified by PCR method,then it was inserted in plasmid pET-21a and transformed into E.coli BL21-Codonplus(DE)3-RIL.The soluble high expressive protein was obtained after induced by 0.1 mmol/LIPTG for 2 h.After ultrasonic and centrifugation,the supernate was heated at 85 ℃ and centrifuged again,then the primary purified Pfu-HSP60 protein was obtained and then it was purified by DEAE Sepharose Fast Flow anion exchange chromatography.After the collected peak was filtered and concentrated,the Pfu-HSP60 protein peak was collected through Sephacryl S-200 gel filtration column.The purity of the recombinant protein Pfu-HSP60 was 94.5% and the total yield was 28.0%.[Conclusion] The research laid the foundation for the study of the structure and function of Pfu-HSP60 protein and the study of the interaction mechanism with other HSP.
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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".