Cloning, sequence analysis, and prokaryotic expression vector construction of glycosyltransferase gene BcUGT8 from Bupleurum chinense
Bibliographic record
Abstract
Objective To clone the full-length cDNA of the uridine diphosphate glycosyltransferase(UGT) gene in Bupleurum chinense(BcUGT8),which may be involved with the saikosaponin biosynthesis,and to construct the prokaryotic expression vector.The work will provide the foundation for its further function verification by in vitro expression and activity analysis of the purified protein.Methods RACE and LD-PCR were used to clone the full-length cDNA of BcUGT8,on the basis of its partial cDNA sequence obtained from our previous high-flux sequencing by Roche(454) GS FLX system.The open reading form(ORF) was PCR cloned using primers with corresponding restriction enzymes cutting sites.The PCR products were digested with corresponding restriction enzymes and then were inserted in expression vector pET-28a(+) to construct the recombinant expression vectors.Results The full-length cDNA of UGT gene was cloned from B.chinense,and the prokaryotic expression vector was obtained.Conclusion The full-length cDNA cloning,sequence analysis,and prokaryotic expression vector construction provide a substantial foundation for follow-up bio-function analysis of BcUGT8 through protein expression,purification,and activity analysis in vitro.
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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) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 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".