Construction of eukaryotic expressing vector pEGFP-N1/PDGF-A for transducting Dermis-derived mesenchymal stem cells
- VernacularTitle:重组真核表达载体pEGFP-N1/PDGF-A的构建及真皮干细胞的转染
- Author:
Guohe YAN
;
Yongping SU
;
Junping WANG
;
Daijie WANG
;
Guoping AI
;
Fengchao WANG
;
Xinze RAN
;
Tianmin CHENG
- Publication Type:Journal Article
- Keywords:
cloning;
eukaryotic expression vector;
platelet-derived growth factor A chain gene;
enhanced green fluorescent protein;
dermis-drived mesenchymal stem cells
- From:Journal of Third Military Medical University
2003;0(20):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone platelet-derived growth factor A chain (PDGF-A) gene and insert PDGF-A gene into. Enhanced green fluorescent protein (EGFP) vector and then transformed into dermis-drived mesenchymal stem cells (DMSCs). Methods cDNA clones encoding human PDGF-A gene were isolated from a human hepatoma cell line mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The PCR amplified fragment of PDGF-A gene was cloned into pMD18-T vector. The eukaryotic expression vector pEGFP-N1/PDGF-A was constructed by subcolone PDGF-A gene into pEGFP-N1 vector. PDGF-A gene was transfected into DMSCs with the help of Fugene 6 transfection reagent. Results Full cDNA sequence encoding human PDGF-A gene had been cloned, which sequence was consistent with the reported sequence in GenBank by sequence assaying. Conclusion cDNA sequence encoding human PDGF-A gene was successfully cloned into pEGFP-N1. The transient expression of PDGF-A gene in DMSCs has been realized.
