Effect of enhanced green fluorescent protein fusion on Ano1 physiological feature.
- Author:
Kai ZHENG
1
;
Hui-Jing XU
1
;
Yu-Xuan ZANG
2
;
Yi-Ju HOU
1
;
Li ZHANG
1
;
Hai-Ou YANG
1
;
Jie ZHU
2
;
Fang FANG
2
;
Feng HAO
3
Author Information
1. Department of Biochemistry Laboratory, Jilin Medical College, Jilin 132013, China.
2. Department of Hematology, Jilin Medical College, Jilin 132013, China.
3. Department of Biochemistry Laboratory, Jilin Medical College, Jilin 132013, China. haof863@126.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Anoctamin-1;
Cell Membrane;
physiology;
Chloride Channels;
metabolism;
Epithelial Cells;
physiology;
Genetic Vectors;
Green Fluorescent Proteins;
metabolism;
Microscopy, Fluorescence;
Plasmids;
Rats;
Recombinant Fusion Proteins;
metabolism;
Thyroid Gland;
cytology;
Transfection
- From:
Acta Physiologica Sinica
2015;67(6):623-628
- CountryChina
- Language:Chinese
-
Abstract:
The aim of the present study was to investigate whether the physiological features of Ano1 were affected by enhanced green fluorescent protein (EGFP) fusing at Ano1 C-terminal. The eukaryotic expression vectors of Ano1 and EGFP-Ano1 were constructed, and these plasmids were transfected into Fischer rat thyroid follicular epithelial (FRT) cells using liposome. The expression and location of Ano1 were examined by using inverted fluorescence microscope. The ability of Ano1 to transport iodide was detected by kinetics experiment of fluorescence quenching. The results showed that both Ano1 and EGFP-Ano1 were expressed on FRT cell membrane and could be activated by Ca(2+). There was no significant difference of the ability to transport iodide between Ano1 and EGFP-Ano1. These results suggest Ano1 and EGFP-Ano1 have similar physiological feature.
