[Cell-penetrating peptide PEP-1-mediated transduction of enhanced green fluorescent protein into human colorectal cancer SW480 cells].
OBJECTIVE
Cell-penetrating peptides, a class of small cationic peptides, could mediate macromolecules transduction into many cell lines. This study was to explore the penetrating ability of PEP-1 in the transduction of enhanced green fluorescent protein (EGFP) into human colorectal cancer cell line SW480.


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. Purified EGFP and PEP-1-EGFP were incubated with SW480 cells. The expression of EGFP in SW480 cells was observed under inverted fluorescent microscope.


RESULTS
EGFP could not be transduced into SW480 cells, whereas PEP-1-EGFP fusion protein could be transduced into SW480 cells and distributed in cytoplasm and nuclei after 1-hour incubation.


CONCLUSIONS
Mediated by PEP-1, EGFP could be transduced efficiently into SW480 cells.
Dong
Xiao
X
Institute of Clinical Medicine, People's Hospital, Yunyang Medical College, Shiyan, Hubei, PR China.
Wang
Jia-Ning
JN
Huang
Yong-Zhang
YZ
Guo
Ling-Yun
LY
Kong
Xia
X
D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D049408:Luminescent Agents / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D049408:Luminescent Agents; C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D003543:Cysteamine