Construction and identification of recombinant adenovirus expressing hPDGF-A and hBD-2
- VernacularTitle:hPDGF-A/hBD_2双基因共表达腺病毒载体的构建及表达
- Author:
Lei HAO
;
Zhongmin ZOU
;
Junping WANG
;
Shiwu DONG
;
Jun DENG
;
Guohe YAN
;
Lianyou WANG
;
Yu NING
;
Dengqun LIU
;
Chengji LUO
;
Yongping SU
- Publication Type:Journal Article
- Keywords:
adenovirus;
PDGF-A;
Defensin ?2;
BMSC
- From:Journal of Third Military Medical University
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To further determine their possible synergistic effect on accelerating wound healing, adenovirus vector containing recombinant human hPDGF-A and hBD2 genes was constructed and the expression of exogenous genes in transformed mesenchymal stem cells derived from rat bone marrow was observed. Methods By putting IRES in the middle of hPDGF-A and hBD2, these two genes were expected to be expressed individually. The shuttle vector was named as pAdTrack-hPDGF-A-IRES2-hBD2, which homologously recombinated with Adeasy-1 in BJ5183 cells and formed the mammalian expression vector pAdeasy-hPDGF-A-IRES2-hBD2. Furthermore, the recombinant vector was packaged in 293 cells into infectious recombinant adenovirus, which were used to infect BMSCs. The expression of hPDGF-A and hBD2 in BMSCs was detected by RT-PCR. Results We successfully constructed recombinant adenovirus vector that simultaneously expressed hPDGF-A and hBD2. The expressions of hPDGF-A and hBD2 were confirmed by RT-PCR on transformed BMSCs. Conclusion The established BMSCs that overexpressed hPDGF-A and hBD2 provide a new strategy of combining cell therapy and gene therapy to promote wound healing, especially the chronic one.
