Construction and expression of TCF4/pcDNA3.0 expression vector
- VernacularTitle:TCF4/pcDNA3.0表达载体的构建及其在毛乳头细胞中的表达
- Author:
Yi LIU
;
Fei HAO
;
Xichuan YANG
- Publication Type:Journal Article
- Keywords:
TCF4;
pcDNA3.0 expression vector;
expression
- From:Journal of Third Military Medical University
1984;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo clone TCF4 (T cell factor 4) gene and construct its eukaryotic expression vector. MethodsThe total RNA was extracted from the aggregated human dermal papilla cells. The full length cDNA encoding TCF4 was obtained by RT- PCR, digested by restriction enzyme, then inserted in the eukaryotic expression vector pcDNA3.0. The sequence and reading frame were confirmed by two restriction enzymes and sequencing. The recombinant vector TCF4/pcDNA3.0 was stably transfected into dermal papilla cells, and the expression changes of TCF4 gene were detected. ResultsTCF4 gene was cloned from dermal papilla cells and its eukaryotic expression vector was constructed. After the identification and sequencing, the reconstructed plasmid was confirmed containing the correct and full nucleotide sequence of TCF4 gene. After stable transfection, the mRNA and protein level of TCF4 gene were up-regulated in dermal papilla cells and the proliferation of dermal papilla cells was promoted. ConclusionThe expression vector TCF4/pcDNA3.0 was constructed successfully and could be expressed in the dermal papilla cells. TCF4 gene can promote the proliferation of the dermal papilla cells.
