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 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) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 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".