Construction and identification of lentiviral vector carrying FOLR1 gene.
- Author:
Mingju HUANG
1
;
Qi WANG
;
Wei ZHANG
;
Qiaoyun YIN
;
Zhuang LI
;
Li LI
Author Information
1. Department of Gynecologic Oncology, Tumor Hospital of Guangxi Medical University, Nanning 530021, China.
- Publication Type:Journal Article
- MeSH:
Cell Line;
Cell Line, Tumor;
Cloning, Molecular;
Female;
Folate Receptor 1;
biosynthesis;
genetics;
Genetic Vectors;
genetics;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Humans;
Kidney;
cytology;
Lentivirus;
genetics;
metabolism;
Ovarian Neoplasms;
pathology;
Polymerase Chain Reaction;
Recombinant Proteins;
biosynthesis;
genetics;
Transfection
- From:
Journal of Biomedical Engineering
2013;30(3):641-650
- CountryChina
- Language:Chinese
-
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.
