Construction of recombinant adenovirus vector carrying human TIMP-1 cDNA and its expression in vitro / 生物医学工程学杂志
Journal of Biomedical Engineering
; (6): 420-424, 2007.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-357685
Responsible library:
WPRO
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.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Recombinant Fusion Proteins
/
Cell Line
/
Adenoviridae
/
Cloning, Molecular
/
DNA, Complementary
/
Carcinoma, Hepatocellular
/
Tissue Inhibitor of Metalloproteinase-1
/
Extracellular Matrix
/
Genetic Vectors
/
Genetics
Limits:
Humans
Language:
Chinese
Journal:
Journal of Biomedical Engineering
Year:
2007
Document type:
Article