Construction of recombinant adenovirus vector carrying human TIMP-1 cDNA and its expression in vitro.
- Author:
Dong XIA
1
;
Lünan YAN
;
Liang XU
;
Yu TONG
;
Huaiquan ZUO
;
Lanying ZHAO
Author Information
1. Department of General Surgery, West China Hospital of Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Carcinoma, Hepatocellular;
metabolism;
Cell Line;
Cloning, Molecular;
DNA, Complementary;
genetics;
Extracellular Matrix;
metabolism;
Genetic Vectors;
Humans;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Tissue Inhibitor of Metalloproteinase-1;
biosynthesis;
genetics
- From:
Journal of Biomedical Engineering
2007;24(2):420-424
- CountryChina
- Language:Chinese
-
Abstract:
The full-length cDNA of hTIMP-1 was obtained from a surgical patient with HCC by the method of RT-PCR. Then it was cloned into the adenoviral shuttle plasmid pAdTrack-CMV, and subsequently cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thereupon, a recombinant adenoviral plasmid containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. The adenoviruses (AdhTIMP-1) were packaged and amplified in adenoviral packaging cells HEK 293. Then the viral titer was checked by green fluorescent protein (GFP), and the expression of hTIMP-1 was detected by the techniques of Western blot and RT-PCR. The recombinant adenovirus vector carrying human TIMP-1 was successfully constructed and expressed in vitro and may pave the way for further application in liver gene therapy.
