Cell-penetrating peptide PEP-1 mediated transmembrane delivery of enhanced green fluorescent protein in vivo of mouse
- VernacularTitle:细胞穿透肽PEP-1介导增强型绿色荧光蛋白在小鼠体内跨膜转导
- Author:
Xiao DONG
;
Jianing WANG
;
Junming TANG
;
Guodong PAN
;
Yongzhang HUANG
;
Jianye YANG
;
Shufen CAO
- Publication Type:Journal Article
- Keywords:
cell-penetrating peptide;
protein transduction domain;
protein transduction;
PEP-1;
enhanced green fluorescent protein
- From:
Basic & Clinical Medicine
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the in vivo transduction capability of fusion protein PEP-1-EGFP with mice.Methods Two prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E.coli BL21(DE3) to express EGFP and fusion protein PEP-1-EGFP,respectively.The expressed EGFP and PEP-1-EGFP were purified with Ni2+-resin affinity chromatography.Five hundred micrograms of EGFP and PEP-1-EGFP fusion protein were injected into mouse through caudal vein,respectively,the mice were euthanized and perfused with PBS 2 hours after administration.Then,the heart,brain,liver,spleen and kidney were removed and sectioned with a cryostat at 7 ?m for visualization with a inverted fluorescent microscope.ResultsThe brain,heart,liver,spleen and kidney injected with PEP-1-EGFP showed bright and homogenous green fluorescence whereas that with EGFP showed no green fluorescence at all.Conclusion The successful expression and purification of PEP-1-EGFP fusion protein and its efficient transduction into mice in vivo provide a basis for the research on transmembrane delivery of macromolecule drugs mediated by the cell-penetrating peptide,PEP-1.
