Construction and identification of lentiviral vector carrying FOLR1 gene / 生物医学工程学杂志
Journal of Biomedical Engineering
; (6): 641-650, 2013.
Article
de Zh
| WPRIM
| ID: wpr-352194
Bibliothèque responsable:
WPRO
ABSTRACT
Through this research a lentiviral vector expressing the gene of folate-binding protein-1 (FOLR1) was constructed and the corrsponding expression products were identified. Firstly, full-length of the FORL1 gene was amplified by PCR and cloned into the plasmid pWPI. Then it was further confirmed by PCR and sequencing. Secondly, after the recombinant pWPI and its helper plasmid co-transfected the virus packaging 293T cells, SKOV3 cells were infected with the virus particles and sorted by flow cytometry. Thirdly, the FOLR1 gene was detected by RT-PCR and its protein expression was detected by Western blot. Finally, the recombinant expression vector was successfully constructed, and lentiviruses were successfully packaged by the 293T cells. A great quantity of green fluorescent cells could be seen after the SKOV3 cells were effectively infected with the lentiviruses carrying the FOLR1 gene. The sorting could be done and detected by cytometrying the FORL1 gene and its stable expression by the two methods above, which laid experimental foundation for exploring its biological function in ovarian cancers.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Tumeurs de l'ovaire
/
Anatomopathologie
/
Protéines recombinantes
/
Transfection
/
Lignée cellulaire
/
Réaction de polymérisation en chaîne
/
Clonage moléculaire
/
Lentivirus
/
Biologie cellulaire
/
Lignée cellulaire tumorale
Type d'étude:
Diagnostic_studies
Limites du sujet:
Female
/
Humans
langue:
Zh
Texte intégral:
Journal of Biomedical Engineering
Année:
2013
Type:
Article