Construction of eukaryotic expression vector of short hairpin RNA for transforming growth factor-beta1.
- Author:
Jing WANG
1
;
Jun-zheng WU
;
Fu-ping GUO
;
Xiu-li ZHU
;
De-sheng WEN
Author Information
- Publication Type:Journal Article
- MeSH: Genetic Vectors; Humans; Immunohistochemistry; Plasmids; RNA, Messenger; RNA, Small Interfering; Transfection; Transforming Growth Factor beta1
- From: West China Journal of Stomatology 2006;24(2):113-116
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the plasmid containing short hairpin RNA (shRNA) of TGF-beta1 expression vector.
METHODSShort chain oligonucleotide was designed according to the TGF-beta1 mRNA sequence provided by Genebank, then DNA segment was gained through annealing after chemosynthesis, and then was cloned to pWH1 vector. The recombinant TGF-beta1 shRNA expression vector was evaluated by using enzyme cutting. At last, the constructed TGF-beta1 expression vector was transfected into salivary gland mucoepidermoid carcinoma (Ms) cells by Lipofectomine TM 2000, and its effect on TGF-beta1 expression was observed by RT-PCR and immunohistochemistry.
RESULTSSuccessful construction was identified by enzyme cutting and the constructed plasmid was called pWH1-TGF-beta1. The shRNA and it inhibited the TGF-beta1 mRNA and protein expression effectively.
CONCLUSIONThe constructed TGF-beta1 shRNA expression vector can block the TGF-beta1 expression in salivary gland mucoepidermoid carcinoma cells.
