The PTD domain of Tat protein enhance GFP protein delivering into myeloma cell SP2/0.
- Author:
Zhi-Qi LI
1
;
Xiao-Bo HU
;
Sheng-Li YANG
;
Yi GONG
Author Information
1. Shanghai Research Center of Biotechnology, Chinese Academy of Sciences, Shanghai 200233, China.
- Publication Type:Journal Article
- MeSH:
Gene Products, tat;
chemistry;
metabolism;
Green Fluorescent Proteins;
Humans;
Luminescent Proteins;
metabolism;
Microscopy, Fluorescence;
Multiple Myeloma;
metabolism;
Protein Transport;
Recombinant Fusion Proteins;
metabolism;
Tumor Cells, Cultured
- From:Chinese Journal of Biotechnology
2002;18(5):644-647
- CountryChina
- Language:Chinese
-
Abstract:
In order to detect the protein delivery mediated by the PTD (protein transduction domain) of TAT Protein, a expression vector, named pT7460-GFP, was constructed by insert the PTD DNA Sequence, followed by a GFP (green fluorescent protein) gene fused in-frame, into the pT7450 vector. The TAT-GFP fusion protein was expressed in the E. coli ER2566. Most of the fusion protein was presented in the inclusion body. The protein was purified by Ni2+ affinity chromatography under denature conditions, then by a Sepharose Q column to remove urea. The soluble denatured protein was added directly to medium containing the Myeloma Cell SP2/0. It came out that the fusion protein could be detected delivered into the cells under fluorescent microscope in a short time.
