Cloning and expression of rat transforming growth factor β1 cDNA in osteoblasts |
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| Authors: | Liu Yong Zheng Qixin Du Jingyuan Zeng Hui Guo Xiaodong Qu Shen |
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| Affiliation: | (1) Department of Orthopaedics, Xiehe Hospital, Tongji Medical University, 430022 Wuhan;(2) Department of Biochemistry, School of Basic Medical Sciences, Tongji Medical University, 430030 Wuhan |
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| Abstract: | Summary Rat transforming growth factor β1 (rTGFβ1) cDNA from rat lymphocytes was cloned by RT-PCR and inserted into pcDNA3 to construct an eukaryotic expression vector, which was named pcDNA3-TGFβ1. The cloned gene was confirmed to code rat TGFβ1 by restriction enzyme analysis. pcDNA3-TGFβ1 plasmid was transfected into rat osteoblasts by using liposome-mediated gene transfer technique and the expression of TGFβ1 was detected by using immunohistochemical staining assay. It was found that the rat TGFβ1 expression product was obviously detectable in the transfected osteoblasts in 48 h. High expression of TGFβ1 was obtained in the rat osteoblasts in which the constructed TGFβ1 expression vector was transfected. |
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| Keywords: | transforming growth factor β 1 gene expression RT-PCR osteoblasts molecular cloning |
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