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Cloning and expression of rat transforming growth factor beta 1 cDNA in osteoblasts
1Department of Orthopaedics, Xiehe Hospital, Tongji Medical University, Wuhan 430022.
Abstract:
Rat transforming growth factor beta 1 (rTGF beta 1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGF beta 1. The cloned gene was confirmed to code rat TGF beta 1 by restriction enzyme analysis. pcDNA3-TGF beta 1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGF beta 1 was detected by using immunohistochemical staining assay. It was found that the rat TGF beta 1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGF beta 1 was obtained in the rat osteoblasts in which the constructed TGF beta 1 expression vector was transfected.