Cloning and expression of rat transforming growth factor beta 1 cDNA in osteoblasts.
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.
Liu
Y
Y
Department of Orthopaedics, Xiehe Hospital, Tongji Medical University, Wuhan 430022.
Zheng
Q
Q
Du
J
J
Zeng
H
H
Guo
X
X
Qu
S
S
D000818:Animals; D000831:Animals, Newborn; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D010006:Osteoblasts / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics*; D053773:Transforming Growth Factor beta1
D018076:DNA, Complementary; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1