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TAT Protein Transduction Peptide Mediated Heterologous Proteins Transduction in C.elegans
WU Yong-hong, SHI Jin-ping, HE Guo-wei, REN Chang-hong, GAO Yan, ZHANG Cheng-gang
China Biotechnology ›› 2011, Vol. 31 ›› Issue (03) : 39-45.
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PDF(794 KB)
TAT Protein Transduction Peptide Mediated Heterologous Proteins Transduction in C.elegans
The TAT protein transduction peptide was enriched in basic amino acids and encoded by the HIV-1 virus. Previous studies have revealed that it could safely and efficiently mediate various heterologous biological macromolecules across a variety of biomembranes, such as the plasmid membrane and the blood-brain barrier et al. To further study its function in mediating heterologous proteins transduction in nematode in vivo, the prokaryotic expression vector pET28b-EGFP and pET28b-TAT-EGFP were constructed and induced by IPTG (final concentration 1 mmol/L), followed with the analysis on the expressed protein by fluorescence microscopy, SDS-PAGE and Western blot. Subsequently, the bacterial cells were coated to the LB medium and directly fed to the nematodes followed with capturing the image at 48 h. Results showed when fed to the nematodes for 48 h, the TAT-EGFP fluorescence signals were clearly distributed in the intestinal cells of the worm, while the EGFP fluorescence signals were mainly distributed in the intestinal cavity of the animal. Furthermore, the cellular morphology of TAT-EGFP had no distinct change compared with the EGFP group and controls. Taken together, the data suggested the TAT protein transduction peptide could mediate heterologous protein expression in C.elegans and provided an alternative approach for development of new drug transporter.
TAT protein transduction peptide / Prokaryotic expression / C.elegans / Transmembrane transduction {{custom_keyword}} /
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