The construction of the recombinant PDGF-BB adenovirus vector and its expression in epidermal stem cells

LI Xin-Beng, ZHANG Ming-Sheng, BAI Li-Meng, QIU Jia-Qi, BAI Wen-Fang, TUN Zhi-Dong, TUN  Bo

Chinese Journal of Clinical Anatomy ›› 2010, Vol. 28 ›› Issue (4) : 434.

Chinese Journal of Clinical Anatomy ›› 2010, Vol. 28 ›› Issue (4) : 434.

The construction of the recombinant PDGF-BB adenovirus vector and its expression in epidermal stem cells

  • LI Xin-ping, ZHANG Ming-sheng, BAI Li-ming, et al.
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Abstract

Objective To construct the recombinant adenovirus vector containing PDGF-BB gene and detect its expression in human epidermal stem cells (hESCs). Methods The core sequence of human PDGF-BB was amplified by PCR from pCMV-SPORT6, and then was cloned to pAd track-CMV. The linearized shuttle plasmid was homogenously recombined with pAdeasy-l in BJ5183, and the potential clone was analyzed by restriction endonuclease digestion. The correct clone was linearized and transfected into HEK293 cells for packing and amplifying so as to obtain adenovirus rAD-PDGF. hESCs were separated from the human prepuce, cultured in vitro, and identified by flow cytometry. hESCs were infected by the harvested virus, and the protein expression of PDGF-BB in hESCs was respectively detected by Western blot. Results PDGF-BB was correctly cloned into the adenovirus vector, and the percentage of putative epidermal stem cells showed expression of the molecular marker was about 88%. The recombinant adenovirus vector containing human PDGF-BB gene was constructed successfully, which could infect hESCs efficiently, with the expression of the PDGF-BB in the infected hESCs detected by Western blotting. Conclusions  The human ESCs infected by constructed adenovirus vector rAD-PDGF could express PDGF-BB successfully.

Key words

Platelet-derived growth factor (PDGE-BB) / Adenovirus / Human epidermal stem cells(hESCs)

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LI Xin-Beng, ZHANG Ming-Sheng, BAI Li-Meng, QIU Jia-Qi, BAI Wen-Fang, TUN Zhi-Dong, TUN  Bo. The construction of the recombinant PDGF-BB adenovirus vector and its expression in epidermal stem cells[J]. Chinese Journal of Clinical Anatomy. 2010, 28(4): 434

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