Construction of the plasmid encoding the fusion protein of hepatocyte growth factor and red fluorescent protein.
- Author:
Ying YANG
1
;
Jun YANG
Author Information
1. Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071, China.
- Publication Type:Journal Article
- MeSH:
Cell Line;
Genetic Vectors;
genetics;
Hepatocyte Growth Factor;
biosynthesis;
genetics;
Humans;
Luminescent Proteins;
biosynthesis;
genetics;
Plasmids;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics
- From:
Journal of Biomedical Engineering
2009;26(6):1286-1290
- CountryChina
- Language:Chinese
-
Abstract:
In order to trace the expression of exogenous hepatocyte growth factor (HGF) in eukaryotic cells exactly, a recombinant plasmid that expresses the fusion protein of HGF and red fluorescent protein (RFP) was constructed. The gene encoding HGF without stop code was amplified from pMD19-T-HGF by PCR technique and then cloned in pDsRed-express-N1. The recombinant plasmid (pDsRed-HGF) was identified by restriction endonuclease enzyme analysis and DNA sequence analysis. pDsRed-HGF was transfected into 293T cells. The expression of HGF mRNA, HGF and red fluorescent protein was detected. The results showed that the target gene sequence in pDsRed-HGF was completely in conformity with HGF cDNA in GenBank, and it was expressed highly in 293T cells. In this study, pDsRed-HGF was successfully constructed and expressed in eukaryotic cells.
