2006•PubMedRequires access
[Construction of eukaryotic expression vector of short hairpin RNA for transforming growth factor-beta1].
Jing Wang, Junzheng Wu, Fuping Guo, Xiu-Li Zhu, De-Sheng Wen
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Abstract
OBJECTIVE: To construct the plasmid containing short hairpin RNA (shRNA) of TGF-beta1 expression vector. METHODS: Short 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. RESULTS: Successful 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. CONCLUSION: The constructed TGF-beta1 shRNA expression vector can block the TGF-beta1 expression in salivary gland mucoepidermoid carcinoma cells.