Construction of the siRNA expression vector and the silencing influence on focal adhesion kinase gene of Tca8113 cell line / 实用口腔医学杂志
Journal of Practical Stomatology
;
(6)2001.
Article
Dans Chinois
| WPRIM
| ID: wpr-545907
ABSTRACT
Objective:
To construct the siRNA expression vector of focal adhesion kinase(FAK) gene and inhibit the expression of FAK gene in tongue cancer cell line Tca8113 by RNA interfering technique.Methods:
According to the encoding sequence of FAK mRNA, two pieces of oligonucleotide sequences were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pGCSilencerTM-U6/Neo siRNA expression vector. After being identified by restriction enzyme method, the recombinant pSilencer-FAK plasmids were transfected into Tca8113 cells. The transfected cells were selected by G418 method. Immuocytochemistry and Western blotting were used to evaluate FAK gene silencing efficiency.Results:
The oligonucleotide fragments were correctly inserted into pGCSilencerTM-U6/Neo vector. FAK expression of the transfected cells was significantly down-regulated by pSilencer-FAK.Conclusion:
The siRNA expression vector of FAK is successfully constructed and FAK expression of Tca8113 cells can be inhibited by RNA interfering technique.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
langue:
Chinois
Texte intégral:
Journal of Practical Stomatology
Année:
2001
Type:
Article
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