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.
Artigo
em Chinês
| WPRIM
| ID: wpr-352194
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Neoplasias Ovarianas
/
Patologia
/
Proteínas Recombinantes
/
Transfecção
/
Linhagem Celular
/
Reação em Cadeia da Polimerase
/
Clonagem Molecular
/
Lentivirus
/
Biologia Celular
/
Linhagem Celular Tumoral
Tipo de estudo:
Estudo diagnóstico
Limite:
Feminino
/
Humanos
Idioma:
Chinês
Revista:
Journal of Biomedical Engineering
Ano de publicação:
2013
Tipo de documento:
Artigo
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