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 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 | 0.001 |
| 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) | 0.001 | 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 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".