Plant pyruvate-dependent gamma-aminobutyrate transaminase: identification of an<i>Arabidopsis</i>cDNA and its expression in<i>Escherichia coli</i>
Bibliographic record
Abstract
Both pyruvate- and 2-oxoglutarate-dependent gamma-aminobutyrate transaminase (GABA-T) activities are present in crude tobacco (Nicotiana tabacum L.) leaf extracts. In this study, GABA:pyruvate-T activity was partially purified using mitochondrial isolation and protein solubilization in 3-[3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, and a combination of chromatographic and electrophoretic procedures. A partial amino acid sequence of the putative 55-kDa GABA-T subunit enabled identification of a predicted Arabidopsis thaliana (L.) Heynh. GABA:pyruvate-T expressed sequence tag and subsequent amplification of a 1515 bp open reading frame encoding a 504-amino acid polypeptide. Computer analysis using web-based tools revealed the presence of a putative mitochondrial signal sequence and a pyridoxal-5-phosphate binding domain in the polypeptide. Functional expression of the GABA-T cDNA in Escherichia coli revealed that the recombinant protein uses pyruvate but not 2-oxoglutarate. The Arabidopsis GABA:pyruvate-T cDNA could form the basis for identification of multiple GABA-T isoforms and generation of GABA-T mutants for determining the fate of GABA nitrogen and elucidating the physiological function of GABA in plants.Key words: amino acceptor, gamma-aminobutyrate, gamma-aminobutyrate transaminase, protein purification, heterologous expression, recombinant protein.
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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 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 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".