Co-expression plasmid construction of vascular endothelial growth factor and Fas ligand and detection of its expression in cultivated cells
Bibliographic record
Abstract
Objective To construct the co expression plasmid of vascular endothelial growth factor (VEGF) and Fas ligand (FasL) and detect its expression in cultivated cells.Methods The primers were designed and synthetized according to the cDNA sequences of human VEGF165 and FasL,then PCR amplication,connection,splicing and insertion were carried out and the recombinant plasmids of pCI VEGF165 and pCI FasL were obtained.The co expression plasmid encoding FasL and VEGF165 named pCI FasL IRES VEGF165 (simplified as pCI FIV) was successfully constructed.The constructed plasmids (2 μg) were transfected to NIH3T3 cell with lipofectamine and the expressions of FasL and VEGF165 were detected by flow cytometry,ELISA and western blot.Results Western blot of the cell splitting solution of pCI VEGF165 and pCI FIV detected the expression of VEGF165 with the estimated molecular weight,and ELISA detection of the supernatant of pCI VEGF165 and pCI FIV revealed a secretory expression of VEGF165.Flow cytometry found the green fluorescent staining cells of pCI FasL and pCI FIV stably transfected NIH3T3 were 98.0% and 96.5% respectively,while that of the empty vector was 3.2%.ELISA detected the soluble FasL expression in the supernatant of pCI FasL and pCI FIV transfected cells.Conclusion The FasL and VEGF co expression plasmid was successfully constructed and it could express secretory VEGF165 and membrane binding FasL and soluble FasL,which lay a foundation for future gene therapy of vascular intimal hyperplasia.
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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".