Construction and expression of the eukaryotic expression vector containing human soluble transforming growth factor beta receptor II.
- Author:
Xiao-hua YANG
1
;
Li-huang ZHANG
;
Li-yun SHI
;
Hang-ping YAO
;
Zi-yang YU
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Animals; Base Sequence; CHO Cells; Cloning, Molecular; Cricetinae; DNA, Complementary; Eukaryotic Cells; metabolism; Genetic Vectors; Humans; Molecular Sequence Data; Plasmids; genetics; Protein-Serine-Threonine Kinases; Receptors, Transforming Growth Factor beta; biosynthesis; genetics; Recombination, Genetic; genetics; Transfection
- From: Journal of Zhejiang University. Medical sciences 2004;33(6):504-508
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a eukaryotic expression vector encoding the gene of extracellular region of type II transforming growth factor beta receptor (sTGFbetaR II), to express the protein in CHO cell line and to determine its biological activity.
METHODSThe extracellular region (amino acids 1-159) of the human TGFbetaR II cDNA was amplified by PCR from a TGFbetaR II chimeric plasmid,and the eukaryotic expression plasmid pCDNA3.1/myc-his(-)B-sTGFbetaR II(pCDNA-sTGFbetaR II) was constructed by inserting the sTGFbetaR II cDNA into the EcoR I/Hind III-digested pCDNA. The DNA sequence was confirmed by double digestion and the pCDNA-sTGFbetaR II plasmid was transfected into the CHO cell line. The sTGFbetaR II protein was confirmed by Western blotting analysis and its biological function was determined.
RESULTSThe specific protein was observed in western blotting, and the protein abrogated the growth-inhibitory effects of TGF-beta1 on mink lung epithelial cells (Mv1Lu).
CONCLUSIONThe eukaryotic expression plasmid pCDNA-sTGFbetaR II has been successfully constructed and the sTGFsTGFbetaR II protein with biological activity obtained.
